The document summarizes Pierre Zwiegers' thesis defense for a Master of Science degree. Zwiegers investigated targeted lentiviral-mediated delivery of progranulin cDNA in a genetic model of amyotrophic lateral sclerosis (ALS). The thesis included background on ALS and the neurotrophic properties of progranulin. Experiments assessed whether early-stage progranulin upregulation could lessen behavioral and neuropathological symptoms in transgenic mSOD1 mice. Results found no effect on disease onset, survival, neuronal loss or inflammation. Discussion addressed limitations of mSOD1 models for clinical translation and need for replicative pre-clinical studies.
Los días 11 y 12 de diciembre de 2014, la Fundación Ramón Areces celebró el Simposio Internacional 'Neuropatías periféricas hereditarias. Desde la biología a la terapéutica' en colaboración con CIBERER-ISCIII y el Centro de Investigación Príncipe Felipe. El tipo más común de estas patologías es la enfermedad de Charcot-Marie-Tooth, un trastorno neuromuscular hereditario con una prevalencia estimada de 17-40 afectados por 100.000 habitantes. Durante estos dos días, investigadores mostraron sus avances en la mejora del diagnóstico y el tratamiento y, por ende, de la aproximación clínica y la calidad de vida de las personas afectadas por estas patologías.
Genes in complex neurological disordersDenise Sheer
The document discusses approaches for identifying genes associated with neurological disorders. It describes several methods: linkage studies, homozygosity mapping, exome and whole genome sequencing, genome-wide association studies, detection of structural variation, and transcriptomics. Each method is summarized, with examples provided of neurological genes identified through various approaches such as exome sequencing. The human genome is also briefly described to provide context. In summary, the document outlines strategies for pinpointing genes underlying neurological conditions.
This document discusses lysosomal biomarkers for neuronal ceroid lipofuscinosis (NCL), a group of rare genetic disorders. The author identifies two key points: 1) Lysosomal changes can provide a rapid method to determine if treatments for NCLs are effective, as these diseases progress slowly and standard clinical methods make efficacy difficult to evaluate. 2) Studying lysosomal changes may provide insights into the functions of deficient NCL proteins and why their deficiency causes disease. The author has identified secondary alterations in lysosomal proteins in mouse models of three NCL types (CLN1, CLN2, CLN3) using mass spectrometry and aims to validate potential biomarkers in cerebrospinal fluid.
This study investigated the effects of neonatal hyperoxia (high oxygen exposure) on neurotrophin gene expression in mice. The researchers found that hyperoxia induced alterations in brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) mRNA expression levels in the isocortex and prefrontal cortex. Specifically, BDNF mRNA expression decreased in the prefrontal cortex but increased in the isocortex, while GDNF mRNA expression increased in both regions following hyperoxia. Hyperoxia also altered mRNA expression levels of DNA methyltransferases DNMT1 and DNMT3a, which regulate neurotrophin gene expression through DNA methylation. These results suggest that neonatal
1) The document summarizes the findings of an international survey of CLN2 experts regarding the natural history, diagnosis and management of CLN2 disease.
2) CLN2 disease is a rare, fatal pediatric neurological disorder caused by TPP1 enzyme deficiency. Initial symptoms typically include seizures, speech/language delays, and developmental regression between 1.5-5 years of age.
3) The survey found delays of 1-4 years between first symptoms and CLN2 diagnosis due to low disease awareness. Timely diagnosis is important to enable future treatment options.
Altered proliferation and networks in neural cells derived from idiopathic au...Masuma Sani
Autism Spectrum Disorders; heterogeneous nature of genetic and brain pathology in ASD– which makes it difficult to produce relevant animal and cell models
The document discusses the cause of Fragile X syndrome, which is a genetic condition associated with intellectual disabilities and certain physical characteristics. There has been some debate around whether deletion or expansion of the CGG trinucleotide in the FMR1 gene causes the syndrome. Some research has found that a substitution of G>C in the CGG trinucleotide leads to gene silencing and causes the syndrome. However, other research from a study of a healthy family found that a CGG>CCG substitution is a non-coding polymorphism and does not affect gene expression or cause the syndrome. The purpose is to analyze which findings are correct regarding the molecular basis of Fragile X syndrome.
Toward Precision Medicine in Neurological Disease by David GoldsteinKnome_Inc
View the webinar at http://www.knome.com/webinar-toward-precision-medicine-neurological-disease. In this presentation, Dr. Goldstein reports progress in identifying pathogenic mutations large-scale scale studies in epilepsy, in particular focusing on identifying de novo mutations as a cause of the epileptic encephalopathies. Next he discusses how sequencing is being used to diagnose rare serious unresolved genetic conditions. Finally, Dr. Goldstein describes a number of examples in which a secure genetic diagnosis has led directly to a change in clinical management.
Los días 11 y 12 de diciembre de 2014, la Fundación Ramón Areces celebró el Simposio Internacional 'Neuropatías periféricas hereditarias. Desde la biología a la terapéutica' en colaboración con CIBERER-ISCIII y el Centro de Investigación Príncipe Felipe. El tipo más común de estas patologías es la enfermedad de Charcot-Marie-Tooth, un trastorno neuromuscular hereditario con una prevalencia estimada de 17-40 afectados por 100.000 habitantes. Durante estos dos días, investigadores mostraron sus avances en la mejora del diagnóstico y el tratamiento y, por ende, de la aproximación clínica y la calidad de vida de las personas afectadas por estas patologías.
Genes in complex neurological disordersDenise Sheer
The document discusses approaches for identifying genes associated with neurological disorders. It describes several methods: linkage studies, homozygosity mapping, exome and whole genome sequencing, genome-wide association studies, detection of structural variation, and transcriptomics. Each method is summarized, with examples provided of neurological genes identified through various approaches such as exome sequencing. The human genome is also briefly described to provide context. In summary, the document outlines strategies for pinpointing genes underlying neurological conditions.
This document discusses lysosomal biomarkers for neuronal ceroid lipofuscinosis (NCL), a group of rare genetic disorders. The author identifies two key points: 1) Lysosomal changes can provide a rapid method to determine if treatments for NCLs are effective, as these diseases progress slowly and standard clinical methods make efficacy difficult to evaluate. 2) Studying lysosomal changes may provide insights into the functions of deficient NCL proteins and why their deficiency causes disease. The author has identified secondary alterations in lysosomal proteins in mouse models of three NCL types (CLN1, CLN2, CLN3) using mass spectrometry and aims to validate potential biomarkers in cerebrospinal fluid.
This study investigated the effects of neonatal hyperoxia (high oxygen exposure) on neurotrophin gene expression in mice. The researchers found that hyperoxia induced alterations in brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) mRNA expression levels in the isocortex and prefrontal cortex. Specifically, BDNF mRNA expression decreased in the prefrontal cortex but increased in the isocortex, while GDNF mRNA expression increased in both regions following hyperoxia. Hyperoxia also altered mRNA expression levels of DNA methyltransferases DNMT1 and DNMT3a, which regulate neurotrophin gene expression through DNA methylation. These results suggest that neonatal
1) The document summarizes the findings of an international survey of CLN2 experts regarding the natural history, diagnosis and management of CLN2 disease.
2) CLN2 disease is a rare, fatal pediatric neurological disorder caused by TPP1 enzyme deficiency. Initial symptoms typically include seizures, speech/language delays, and developmental regression between 1.5-5 years of age.
3) The survey found delays of 1-4 years between first symptoms and CLN2 diagnosis due to low disease awareness. Timely diagnosis is important to enable future treatment options.
Altered proliferation and networks in neural cells derived from idiopathic au...Masuma Sani
Autism Spectrum Disorders; heterogeneous nature of genetic and brain pathology in ASD– which makes it difficult to produce relevant animal and cell models
The document discusses the cause of Fragile X syndrome, which is a genetic condition associated with intellectual disabilities and certain physical characteristics. There has been some debate around whether deletion or expansion of the CGG trinucleotide in the FMR1 gene causes the syndrome. Some research has found that a substitution of G>C in the CGG trinucleotide leads to gene silencing and causes the syndrome. However, other research from a study of a healthy family found that a CGG>CCG substitution is a non-coding polymorphism and does not affect gene expression or cause the syndrome. The purpose is to analyze which findings are correct regarding the molecular basis of Fragile X syndrome.
Toward Precision Medicine in Neurological Disease by David GoldsteinKnome_Inc
View the webinar at http://www.knome.com/webinar-toward-precision-medicine-neurological-disease. In this presentation, Dr. Goldstein reports progress in identifying pathogenic mutations large-scale scale studies in epilepsy, in particular focusing on identifying de novo mutations as a cause of the epileptic encephalopathies. Next he discusses how sequencing is being used to diagnose rare serious unresolved genetic conditions. Finally, Dr. Goldstein describes a number of examples in which a secure genetic diagnosis has led directly to a change in clinical management.
Gene therapy involves introducing normal genes into patients to compensate for mutated genes that cause disease. The first gene therapy trial treated a girl with severe combined immunodeficiency. While it initially strengthened her immune system, the effects only lasted a few months. Gene therapy shows promise for diseases caused by single gene defects like cystic fibrosis, but faces challenges like short-lived effects, immune responses, and safety issues. Continued research aims to address these challenges through techniques like RNA interference and improved gene delivery methods.
Gene therapy involves inserting normal genes into cells to treat genetic diseases. The first successful gene therapy trial treated a patient with severe combined immunodeficiency (SCID) caused by adenosine deaminase (ADA) deficiency. The procedure involved removing the patient's lymphocytes, infecting them with a retrovirus carrying the normal ADA gene, allowing the gene to integrate and express ADA, and returning the modified lymphocytes to the patient to permanently produce the enzyme. This landmark trial established gene therapy as a viable treatment approach for genetic diseases.
Gene therapy is the science of making specific changes to the human genome to improve it or achieve therapeutic effects for gene-related diseases. This document discusses gene therapy in three sentences: it defines gene therapy, describes the three main types (germline, somatic, vaccination), and explains that somatic gene therapy aims to treat body cells without affecting future generations. The document then discusses the process of gene therapy including identifying defective genes, creating a replacement gene, and delivering the new gene into the body.
Gene therapy is an experimental technique that uses genes to treat disease by inserting genes into patients' cells instead of using drugs or surgery. Recent research has shown promising advances in gene therapy to treat various diseases. However, gene therapy still faces technical challenges such as safely delivering genes to target cells and tissues, potential immune responses, and difficulty treating complex multigenic disorders. While still experimental, gene therapy offers hope for treating currently incurable conditions.
This document provides an overview of gene therapy including its principles, approaches, development, types, vectors, delivery methods, examples, advantages, and disadvantages. Gene therapy involves inserting genes into cells to treat diseases caused by defective genes. There are two main approaches - germline gene therapy, which alters the germ cells and is passed to offspring, and somatic gene therapy, which alters non-reproductive cells only in the individual. Gene therapy development involves pre-clinical and clinical trials. Vectors like viruses are used to deliver therapeutic genes. Examples include treating severe combined immunodeficiency. While gene therapy has potential benefits, there are also risks like immune responses and ensuring genes reach the right cells.
From “Artificial” to “Real”: What 24/7 Home Cage Monitoring Teaches Us In Pre...InsideScientific
In this webinar Dr. Stefano Gaburro, Scientific Director at Tecniplast, will present an innovative non-invasive and scalable technique called Digital Ventilated Cage (DVC) that is meant to perform longitudinal studies for neurodegenerative disease models using long term monitoring of mice in a stress-free environment.
In the second part of the webinar, Dr. Brun Ulfhake from Karolinska Institutet will show how this technique can be used to study biorhythmicity (circadian and circannual) of small rodents and approaches to characterize and extract metrics of the spontaneous home-cage way of life for mice. These metrics may translate better to behavioral observations made in humans.
Dougherty Reeves Lucas Gamble Lesort Cowell 2012Elizabeth Lucas
This study investigated Purkinje cell dysfunction in an animal model of Huntington's disease (HD). The study found:
1) Reductions in expression of calcium-binding proteins and GABA enzymes in the cerebellum of HD model mice, specifically in the Purkinje cell layer, as shown by gene expression analysis and immunohistochemistry.
2) No loss of Purkinje cells in presymptomatic HD model mice, but significant Purkinje cell loss by end-stage, as shown by stereological analysis.
3) Impaired Purkinje cell firing in presymptomatic HD model mice, prior to accumulation of huntingtin protein and cell loss, indicating early Purkinje cell
This document discusses advances in gene therapy. It begins by explaining what gene therapy is and provides examples of how it has been used to treat children with severe immune deficiencies who previously had to live in protective bubbles. While bone marrow transplants can cure these conditions, they often fail due to graft-versus-host disease. Gene therapy offers advantages over bone marrow transplants by using a patient's own cells with corrected genes, avoiding this complication. The document outlines the process of doing gene therapy outside the body by inserting genes into viruses to deliver them to patients' cells. It notes some of the early successes and challenges of gene therapy clinical trials.
Bioinformatic data analysis – comparison from three human studies using diffe...Agnieszka Caruso
This document summarizes three bioinformatics studies comparing gene expression data from human samples using different Affymetrix platforms: 1) comparing periodontal ligament and gingival cells, 2) identifying genes regulated by nuclear IGFBP5 in osteoblasts, and 3) effects of LPS on osteoblasts. Key findings include identification of differentially expressed genes and biological processes between tissue types in study 1, and processes related to cell cycle, proliferation and splicing regulated by IGFBP5 in study 2. Study 3 on LPS effects found upregulation of factors stimulating osteoclasts and downregulation of processes like mitosis and cell cycle. The document discusses analysis tools and validation of array results.
The document discusses gene therapy, including:
1. Defining genes and discussing early milestones in gene isolation and engineering.
2. Explaining the goal of gene therapy to introduce normal genes to compensate for defective genes.
3. Describing various methods of gene delivery including viral and non-viral vectors.
4. Discussing challenges in targeting specific cells/tissues and ensuring safe expression levels.
Genome Editing & Gene Therapy by Eric KelsicImpact.Tech
Slides from the Genome editing & gene therapy Impact.tech seminar, hosted by Eric Kelsic on June 11th, 2019.
The seminar covers the experiments and inventions that led to the development of genome editing technologies. These inventions were derived from life itself: isolated from natural organisms and adapted for scientific and therapeutic goals. You will learn the history of how genome engineering tools, including CRISPR, and delivery technology, including AAV capsids, were created in their modern form. The seminar explores how genome editing and gene therapy technologies are giving individuals control over their own genomes, focusing on the treatment of genetic diseases. It will describe major companies and emerging trends in the gene therapy industry. Finally, the seminar will discuss how and where new discoveries, including accelerated algorithms for genetic engineering, will lead us in the near and distant future.
Eric Kelsic, PhD, is the founder and CEO of Dyno Therapeutics, a VC-backed biotech located in Cambridge, Massachusetts. Dyno is leading a machine learning revolution to develop enhanced capsid proteins that enable new gene and genome editing therapies. Eric co-developed the technology underlying Dyno’s machine-guided protein engineering platform as a Staff Scientist in George Church’s lab at the Wyss Institute of Harvard Medical School. He holds a PhD in Systems Biology from Harvard University and a BS in Physics from Caltech.
This study tested intrathecal enzyme replacement therapy (ERT) in a mouse model of infantile Batten disease. Mice lacking the enzyme palmitoyl-protein thioesterase 1 (PPT1) received a single intrathecal injection of recombinant human PPT1 at 6 weeks of age. This prevented decline in motor function, improved survival, and reduced brain and spinal cord pathology compared to untreated mice. The effects were similar to ERT for another form of Batten disease. This suggests intrathecal ERT may help treat infantile Batten disease caused by PPT1 deficiency in humans.
This document provides an overview of gene therapy, including types, approaches, vectors used, methods of delivery, advantages, disadvantages, applications, and recent advances. It discusses somatic cell gene therapy, which aims to correct genetic defects in non-reproductive cells, and germline gene therapy, which could pass alterations to future generations but poses more risks. Ex vivo and in vivo gene therapy approaches are described. Viral and non-viral vectors as well as various delivery methods are outlined. Some applications including cystic fibrosis and cancer are highlighted. Risks and ethical considerations are also mentioned.
Thyroxin regulates bdnf expression to promote survival ofSmawi GH
This study examined the effects of thyroxin on brain-derived neurotrophic factor (BDNF) expression and neuronal survival and regeneration following injury in rat hippocampal slices. The results showed that thyroxin promoted survival of injured neurons by upregulating BDNF expression and downregulating the potassium-chloride cotransporter KCC2. Thyroxin also promoted regeneration of injured neurons by increasing axonal growth across lesions. These effects suggest thyroxin could be a potential therapeutic agent for treating central nervous system injuries by mimicking developmental processes.
The future of treatments for PD psych research symposium poster 1_20162Morgan Stafford
This document provides information on Parkinson's disease (PD) and current research. It discusses:
- LRRK2 gene mutations as a common genetic cause of PD and how they disrupt protein recycling and cause cell death.
- Current treatments for PD like drugs that supplement dopamine or inhibit enzymes that break it down, but these only treat symptoms and have side effects.
- Promising research on closed-loop deep brain stimulation and cell transplant therapies that could modify disease progression.
- Ongoing clinical studies of these future treatments and the hope they provide for improving PD treatment.
Gene therapy aims to treat Parkinson's disease by inserting genes into cells to correct deficiencies. It has advantages over traditional treatments by potentially targeting the underlying causes and slowing disease progression. Two main approaches are ex vivo gene therapy, where cells are modified outside the body, and in vivo gene therapy through direct brain injection. Viral vectors are commonly used to deliver genes but carry risks. Ongoing research is exploring using gene therapy to restore dopamine synthesis, modulate basal ganglia activity, or provide neuroprotection for Parkinson's.
Gene therapy involves inserting a normal gene into cells to compensate for a defective gene that causes disease. There are four main approaches: gene replacement, gene repair, gene regulation, and immunization. The first human gene therapy trial took place in 1990 for severe combined immunodeficiency, but it only worked temporarily. While progress is being made, challenges remain such as immune responses, short-lived effects, and difficulties targeting multi-gene disorders.
Role of Brain Derived Neurotrophic Factor (BDNF) in NEURODEVELOPMENTWasiu Adeseji
This document discusses brain-derived neurotrophic factor (BDNF), a protein important for brain development and function. It describes how BDNF is encoded by a gene on chromosome 11 and binds to two receptors, TrkB and p75. The document outlines BDNF's roles in cell survival, differentiation, migration and development. It also discusses BDNF's importance for learning, memory and synaptic plasticity, and implications in neurodegenerative diseases and neurogenesis.
Genome sequencing uncovers MS phenocopies in primary progressive multiple scl...Sherman Jia
This document summarizes a study that performed whole-genome sequencing on patients with primary progressive multiple sclerosis (PPMS) and controls. The key findings were:
1) Some PPMS patients carried rare mutations in genes associated with neurological disorders that resemble MS, called phenocopies.
2) PPMS patients were enriched for rare mutations in genes that cause spastic paraplegias, a type of motor neuron disorder. This enrichment was unique to PPMS and not seen in relapsing-remitting MS.
3) Additional research is needed to understand how both rare and common genetic variations contribute to MS pathogenesis. The findings suggest PPMS may have features of both a complex genetic disease and monogenic
Next Generation Epigenetic Profiling for Environmental Health Sciences
The document discusses next generation epigenetic profiling technologies and their applications in environmental health sciences. It provides an overview of epigenetics and biology, next generation epigenetic profiling technologies like methylation arrays and sequencing, and applications in areas like nutrition, circadian rhythms, temperature stress, and small molecules. The technologies allow genome-wide analysis of epigenetic marks at higher resolution and lower cost than previous methods. This enables studying how environmental exposures may influence human health by altering epigenetic regulation of genes.
Visit this site http://lvnvfundingllc.org/ for more information on LVNV Funding LLC. By going through an intermediary such as collection agencies and attorneys, LVNV Funding LLC tries to side step any liability to comply with the principal purpose of the Fair Debt Collection Practices Act.
This document appears to be a review of related literature on the topic of Telos but provides no actual information. It contains only the title "REVIEW OF RELATED LITERATURE" followed by the word "TELOS" with no other text.
Gene therapy involves introducing normal genes into patients to compensate for mutated genes that cause disease. The first gene therapy trial treated a girl with severe combined immunodeficiency. While it initially strengthened her immune system, the effects only lasted a few months. Gene therapy shows promise for diseases caused by single gene defects like cystic fibrosis, but faces challenges like short-lived effects, immune responses, and safety issues. Continued research aims to address these challenges through techniques like RNA interference and improved gene delivery methods.
Gene therapy involves inserting normal genes into cells to treat genetic diseases. The first successful gene therapy trial treated a patient with severe combined immunodeficiency (SCID) caused by adenosine deaminase (ADA) deficiency. The procedure involved removing the patient's lymphocytes, infecting them with a retrovirus carrying the normal ADA gene, allowing the gene to integrate and express ADA, and returning the modified lymphocytes to the patient to permanently produce the enzyme. This landmark trial established gene therapy as a viable treatment approach for genetic diseases.
Gene therapy is the science of making specific changes to the human genome to improve it or achieve therapeutic effects for gene-related diseases. This document discusses gene therapy in three sentences: it defines gene therapy, describes the three main types (germline, somatic, vaccination), and explains that somatic gene therapy aims to treat body cells without affecting future generations. The document then discusses the process of gene therapy including identifying defective genes, creating a replacement gene, and delivering the new gene into the body.
Gene therapy is an experimental technique that uses genes to treat disease by inserting genes into patients' cells instead of using drugs or surgery. Recent research has shown promising advances in gene therapy to treat various diseases. However, gene therapy still faces technical challenges such as safely delivering genes to target cells and tissues, potential immune responses, and difficulty treating complex multigenic disorders. While still experimental, gene therapy offers hope for treating currently incurable conditions.
This document provides an overview of gene therapy including its principles, approaches, development, types, vectors, delivery methods, examples, advantages, and disadvantages. Gene therapy involves inserting genes into cells to treat diseases caused by defective genes. There are two main approaches - germline gene therapy, which alters the germ cells and is passed to offspring, and somatic gene therapy, which alters non-reproductive cells only in the individual. Gene therapy development involves pre-clinical and clinical trials. Vectors like viruses are used to deliver therapeutic genes. Examples include treating severe combined immunodeficiency. While gene therapy has potential benefits, there are also risks like immune responses and ensuring genes reach the right cells.
From “Artificial” to “Real”: What 24/7 Home Cage Monitoring Teaches Us In Pre...InsideScientific
In this webinar Dr. Stefano Gaburro, Scientific Director at Tecniplast, will present an innovative non-invasive and scalable technique called Digital Ventilated Cage (DVC) that is meant to perform longitudinal studies for neurodegenerative disease models using long term monitoring of mice in a stress-free environment.
In the second part of the webinar, Dr. Brun Ulfhake from Karolinska Institutet will show how this technique can be used to study biorhythmicity (circadian and circannual) of small rodents and approaches to characterize and extract metrics of the spontaneous home-cage way of life for mice. These metrics may translate better to behavioral observations made in humans.
Dougherty Reeves Lucas Gamble Lesort Cowell 2012Elizabeth Lucas
This study investigated Purkinje cell dysfunction in an animal model of Huntington's disease (HD). The study found:
1) Reductions in expression of calcium-binding proteins and GABA enzymes in the cerebellum of HD model mice, specifically in the Purkinje cell layer, as shown by gene expression analysis and immunohistochemistry.
2) No loss of Purkinje cells in presymptomatic HD model mice, but significant Purkinje cell loss by end-stage, as shown by stereological analysis.
3) Impaired Purkinje cell firing in presymptomatic HD model mice, prior to accumulation of huntingtin protein and cell loss, indicating early Purkinje cell
This document discusses advances in gene therapy. It begins by explaining what gene therapy is and provides examples of how it has been used to treat children with severe immune deficiencies who previously had to live in protective bubbles. While bone marrow transplants can cure these conditions, they often fail due to graft-versus-host disease. Gene therapy offers advantages over bone marrow transplants by using a patient's own cells with corrected genes, avoiding this complication. The document outlines the process of doing gene therapy outside the body by inserting genes into viruses to deliver them to patients' cells. It notes some of the early successes and challenges of gene therapy clinical trials.
Bioinformatic data analysis – comparison from three human studies using diffe...Agnieszka Caruso
This document summarizes three bioinformatics studies comparing gene expression data from human samples using different Affymetrix platforms: 1) comparing periodontal ligament and gingival cells, 2) identifying genes regulated by nuclear IGFBP5 in osteoblasts, and 3) effects of LPS on osteoblasts. Key findings include identification of differentially expressed genes and biological processes between tissue types in study 1, and processes related to cell cycle, proliferation and splicing regulated by IGFBP5 in study 2. Study 3 on LPS effects found upregulation of factors stimulating osteoclasts and downregulation of processes like mitosis and cell cycle. The document discusses analysis tools and validation of array results.
The document discusses gene therapy, including:
1. Defining genes and discussing early milestones in gene isolation and engineering.
2. Explaining the goal of gene therapy to introduce normal genes to compensate for defective genes.
3. Describing various methods of gene delivery including viral and non-viral vectors.
4. Discussing challenges in targeting specific cells/tissues and ensuring safe expression levels.
Genome Editing & Gene Therapy by Eric KelsicImpact.Tech
Slides from the Genome editing & gene therapy Impact.tech seminar, hosted by Eric Kelsic on June 11th, 2019.
The seminar covers the experiments and inventions that led to the development of genome editing technologies. These inventions were derived from life itself: isolated from natural organisms and adapted for scientific and therapeutic goals. You will learn the history of how genome engineering tools, including CRISPR, and delivery technology, including AAV capsids, were created in their modern form. The seminar explores how genome editing and gene therapy technologies are giving individuals control over their own genomes, focusing on the treatment of genetic diseases. It will describe major companies and emerging trends in the gene therapy industry. Finally, the seminar will discuss how and where new discoveries, including accelerated algorithms for genetic engineering, will lead us in the near and distant future.
Eric Kelsic, PhD, is the founder and CEO of Dyno Therapeutics, a VC-backed biotech located in Cambridge, Massachusetts. Dyno is leading a machine learning revolution to develop enhanced capsid proteins that enable new gene and genome editing therapies. Eric co-developed the technology underlying Dyno’s machine-guided protein engineering platform as a Staff Scientist in George Church’s lab at the Wyss Institute of Harvard Medical School. He holds a PhD in Systems Biology from Harvard University and a BS in Physics from Caltech.
This study tested intrathecal enzyme replacement therapy (ERT) in a mouse model of infantile Batten disease. Mice lacking the enzyme palmitoyl-protein thioesterase 1 (PPT1) received a single intrathecal injection of recombinant human PPT1 at 6 weeks of age. This prevented decline in motor function, improved survival, and reduced brain and spinal cord pathology compared to untreated mice. The effects were similar to ERT for another form of Batten disease. This suggests intrathecal ERT may help treat infantile Batten disease caused by PPT1 deficiency in humans.
This document provides an overview of gene therapy, including types, approaches, vectors used, methods of delivery, advantages, disadvantages, applications, and recent advances. It discusses somatic cell gene therapy, which aims to correct genetic defects in non-reproductive cells, and germline gene therapy, which could pass alterations to future generations but poses more risks. Ex vivo and in vivo gene therapy approaches are described. Viral and non-viral vectors as well as various delivery methods are outlined. Some applications including cystic fibrosis and cancer are highlighted. Risks and ethical considerations are also mentioned.
Thyroxin regulates bdnf expression to promote survival ofSmawi GH
This study examined the effects of thyroxin on brain-derived neurotrophic factor (BDNF) expression and neuronal survival and regeneration following injury in rat hippocampal slices. The results showed that thyroxin promoted survival of injured neurons by upregulating BDNF expression and downregulating the potassium-chloride cotransporter KCC2. Thyroxin also promoted regeneration of injured neurons by increasing axonal growth across lesions. These effects suggest thyroxin could be a potential therapeutic agent for treating central nervous system injuries by mimicking developmental processes.
The future of treatments for PD psych research symposium poster 1_20162Morgan Stafford
This document provides information on Parkinson's disease (PD) and current research. It discusses:
- LRRK2 gene mutations as a common genetic cause of PD and how they disrupt protein recycling and cause cell death.
- Current treatments for PD like drugs that supplement dopamine or inhibit enzymes that break it down, but these only treat symptoms and have side effects.
- Promising research on closed-loop deep brain stimulation and cell transplant therapies that could modify disease progression.
- Ongoing clinical studies of these future treatments and the hope they provide for improving PD treatment.
Gene therapy aims to treat Parkinson's disease by inserting genes into cells to correct deficiencies. It has advantages over traditional treatments by potentially targeting the underlying causes and slowing disease progression. Two main approaches are ex vivo gene therapy, where cells are modified outside the body, and in vivo gene therapy through direct brain injection. Viral vectors are commonly used to deliver genes but carry risks. Ongoing research is exploring using gene therapy to restore dopamine synthesis, modulate basal ganglia activity, or provide neuroprotection for Parkinson's.
Gene therapy involves inserting a normal gene into cells to compensate for a defective gene that causes disease. There are four main approaches: gene replacement, gene repair, gene regulation, and immunization. The first human gene therapy trial took place in 1990 for severe combined immunodeficiency, but it only worked temporarily. While progress is being made, challenges remain such as immune responses, short-lived effects, and difficulties targeting multi-gene disorders.
Role of Brain Derived Neurotrophic Factor (BDNF) in NEURODEVELOPMENTWasiu Adeseji
This document discusses brain-derived neurotrophic factor (BDNF), a protein important for brain development and function. It describes how BDNF is encoded by a gene on chromosome 11 and binds to two receptors, TrkB and p75. The document outlines BDNF's roles in cell survival, differentiation, migration and development. It also discusses BDNF's importance for learning, memory and synaptic plasticity, and implications in neurodegenerative diseases and neurogenesis.
Genome sequencing uncovers MS phenocopies in primary progressive multiple scl...Sherman Jia
This document summarizes a study that performed whole-genome sequencing on patients with primary progressive multiple sclerosis (PPMS) and controls. The key findings were:
1) Some PPMS patients carried rare mutations in genes associated with neurological disorders that resemble MS, called phenocopies.
2) PPMS patients were enriched for rare mutations in genes that cause spastic paraplegias, a type of motor neuron disorder. This enrichment was unique to PPMS and not seen in relapsing-remitting MS.
3) Additional research is needed to understand how both rare and common genetic variations contribute to MS pathogenesis. The findings suggest PPMS may have features of both a complex genetic disease and monogenic
Next Generation Epigenetic Profiling for Environmental Health Sciences
The document discusses next generation epigenetic profiling technologies and their applications in environmental health sciences. It provides an overview of epigenetics and biology, next generation epigenetic profiling technologies like methylation arrays and sequencing, and applications in areas like nutrition, circadian rhythms, temperature stress, and small molecules. The technologies allow genome-wide analysis of epigenetic marks at higher resolution and lower cost than previous methods. This enables studying how environmental exposures may influence human health by altering epigenetic regulation of genes.
Visit this site http://lvnvfundingllc.org/ for more information on LVNV Funding LLC. By going through an intermediary such as collection agencies and attorneys, LVNV Funding LLC tries to side step any liability to comply with the principal purpose of the Fair Debt Collection Practices Act.
This document appears to be a review of related literature on the topic of Telos but provides no actual information. It contains only the title "REVIEW OF RELATED LITERATURE" followed by the word "TELOS" with no other text.
Renovate Your Retrospective - Adam England - KCDC 2015Adam England
Presentation from Kansas City Developer Conference, June 25, 2015. Adam England discusses common problems in agile retrospectives, and provides solutions for getting to the root cause, and making your retrospectives fun again.
development and experimental validation of a global simulation model of an high power electrical drive (Ansaldo Sistemi Industiali) in a cement plant (Italcementi)
The document discusses reviewing related literature and studies for research. It describes conceptual literature as non-empirical material from various sources that can provide ideas for research problems and theoretical frameworks. Research literature refers to empirical studies from published and unpublished local and foreign sources that can avoid duplication and guide research design. A thorough review of related literature and studies is important as it helps identify gaps, compare variables, and establish trends to strengthen a study. The document provides examples of sources to review and guidelines for writing the literature review introduction and citing sources.
Literature Review (Review of Related Literature - Research Methodology)Dilip Barad
Literature Review or Review of Related Literature is one of the most vital stages in any research. This presentation attempts to throw some light on the process and important aspects of literature review.
Molecular techniques for pathology research - MDX .pdfsabyabby
This document discusses molecular techniques used in pathology research such as PCR, microarrays, next generation sequencing, immunohistochemistry, ELISA, and Western blotting. It provides details on each technique including the basic principles, applications in research, and examples of uses in studies of gene expression, cancer, bone disease, and growth retardation. The learning outcomes are to understand these techniques and their uses in basic and clinical research.
This document summarizes research on Tuberous Sclerosis Complex (TSC), including key facts about the disease, progress that has been made in understanding the genetics and molecular mechanisms, development of treatments like Everolimus, and ongoing areas of research focus like clinical trials of new drugs and studying disease mechanisms using cellular and animal models.
This document outlines Paraneoplastic Neurological Disorder (PND), including its history, definition, pathophysiology, clinical features, diagnosis criteria, treatment, and recommendations. PND is a neurological dysfunction associated with but not directly caused by cancer. It can affect the central, peripheral, or autonomic nervous systems. Onconeural antibodies are present in 60-70% of cases and help diagnose PND. Common PNDs include limbic encephalitis, cerebellar degeneration, and opsoclonus-myoclonus. Treatment involves treating the underlying cancer, immunotherapy like steroids, IVIg, and plasma exchange, and symptom management. Patients with suspected PND should be regularly
This document outlines Paraneoplastic Neurological Disorder (PND), including its history, definition, pathophysiology, clinical features, diagnosis criteria, treatment, and recommendations. PND is a neurological dysfunction associated with but not directly caused by cancer. It can affect the central, peripheral, or autonomic nervous systems. Onconeural antibodies are present in 60-70% of cases and help diagnose PND. Common PNDs include limbic encephalitis, cerebellar degeneration, and opsoclonus-myoclonus. Treatment involves treating the underlying cancer, immunotherapy like steroids or IVIg, and symptom management. Patients with suspected PND should be regularly screened for cancer for
The future: Presentation by Gavin GiovannoniMS Trust
This document summarizes information from a presentation on multiple sclerosis (MS). It begins with disclosures from the presenter regarding compensation received from pharmaceutical companies. It then provides images and diagrams on various topics related to MS, including: cortical lesions; brain atrophy across disease stages; remyelination pathways and targets like LINGO-1; and study designs testing potential neuroprotective and remyelinating agents. One study examines the drug phenytoin in acute optic neuritis to assess neuroprotection by measuring retinal nerve fiber layer thickness. In summary, the document reviews MS pathology and potential new therapeutic strategies targeting remyelination and neuroprotection that are being investigated in clinical trials.
Among patients with relapsing-remitting multiple sclerosis (MS) who underwent nonmyeloablative hematopoietic stem cell transplantation (HSCT), the following results were observed:
1) Scores on the Expanded Disability Status Scale (EDSS) and Neurologic Rating Scale (NRS) improved significantly from pre-transplant levels at 2-year and 4-year follow-ups, indicating reduced neurological disability.
2) Fifty percent of patients showed at least a 1-point improvement on the EDSS at 2 years, increasing to 64% at 4 years.
3) Secondary outcomes including quality of life, walking ability, and lesion volume also significantly improved from pre-transplant
Creutzfeld-Jakob Disease: Diagnosis and Management of Prion Diseasesapplebyb
I. Dr. Brian Appleby discusses diagnosing and managing Creutzfeldt-Jakob disease (CJD), including using EEG, MRI, CSF tests and brain biopsy for diagnosis. Symptomatic treatments aim to improve quality of life.
II. Risk of CJD transmission in clinical settings is low with standard precautions. Care involves managing symptoms, supporting patients and caregivers, and facilitating end-of-life planning.
III. Current research includes studies of bioassays, diagnostic factors, and treatments like art therapy, with the goal of better understanding and caring for those with prion diseases.
Prions in the Urine of Patients with Variant Creutzfeldt–Jakob Diseasedmarquezobando
This document summarizes a study investigating the presence of prions in urine samples from patients with variant Creutzfeldt-Jakob disease (vCJD). The study found that the prion protein PrPSc was detected in the urine of 13 out of 14 vCJD patients using the protein misfolding cyclic amplification (PMCA) technique. Concentrations of PrPSc in urine were estimated to be around 1x10-16 grams per milliliter. The ability to detect PrPSc in urine suggests it may be a useful biomarker for diagnosing vCJD.
1) Clinical trials in progressive multiple sclerosis (MS) face many challenges due to heterogeneity of the condition and unclear pathological mechanisms.
2) Current trials are exploring therapies targeting inflammation, neuroprotection, and repair, with mixed results. Adaptive trial designs may help overcome challenges.
3) Future research priorities include better defining MS phenotypes, identifying pathological drivers of progression, and developing biomarkers to aid trial design and measure treatment response. Several promising therapies are now in late-stage clinical testing.
Temporal-Spatial Expressions of Spy1 in Rat Sciatic Nerve After CrushJiao Yang
1. The study examined the expression of the cell cycle protein Spy1 in a rat sciatic nerve crush injury model over time.
2. Spy1 expression was found to gradually increase after injury, peaking at day 3, due to increased expression in both axons and Schwann cells.
3. Spy1 expression correlated with Schwann cell proliferation after injury and Spy1 was found to localize in axons in the injured segment but did not co-localize with the growth protein GAP43.
Open Source Pharma /Genomics and clinical practice / Prof Hosur opensourcepharmafound
Genomics is increasingly being used in clinical practice to inform diagnosis and treatment. The document discusses several examples where genomics has identified disease-causing genes and mutations, leading to new treatments. It also describes methods for genome sequencing and variant calling. The author's studies on epilepsy found variants in genes related to brain function in drug-resistant epilepsy patients. A large number of variants were in the 3' UTR, suggesting expression level variations in these genes may cause epilepsy.
My presentation delivered at the MS Symposium of the Jewish Hospital Berlin (https://www.juedisches-krankenhaus.de/home.html) held on 29 Nar 2023 at the Centrum Judaicum, Oranienburger Strasse, Berlin
4-1. Steroid-sensitive nephrotic syndrome. Francesco Emma (eng)KidneyOrgRu
This document provides definitions and guidelines for treating steroid-sensitive nephrotic syndrome (SSNS) in children. It discusses that SSNS responds well to prednisone treatment within 4 weeks. The optimal duration of prednisone treatment is debated, with some evidence that longer treatment (3-6 months) reduces relapse risk compared to shorter courses, though it increases steroid toxicity risks. For children who frequently relapse or are steroid dependent, calcineurin inhibitors, mycophenolate mofetil, levamisole, and rituximab may be effective steroid-sparing agents, with varying risks and levels of evidence supporting each option. The document outlines guidelines but notes treatment must be tailored to each individual child
This document discusses the role of functional imaging like PET and SPECT in evaluating patients for epilepsy surgery. It provides information on:
1) When epilepsy surgery evaluation is performed for drug-resistant epilepsy cases. PET and SPECT can help identify the seizure focus when MRI and EEG findings are inconclusive.
2) PET typically shows hypometabolism in the seizure focus interictally and hypermetabolism ictally and postictally. Specific PET tracers can provide data on neurotransmitter systems that may be involved.
3) PET and SPECT findings like hypometabolism or hypoperfusion in the seizure focus can help lateralize and localize the epileptogenic zone to
BIOMARKERS AND SCHIZOPHRENIA1233445677.pptRobinBaghla
Biomarkers are objective indicators of disease that can aid in diagnosis. For schizophrenia, potential biomarkers studied include neurochemical, genetic, imaging, and neurophysiological factors. Dopamine and glutamate dysfunction are implicated. Imaging shows reductions in prefrontal and temporal gray matter volume. Neurophysiological measures like P50 sensory gating and P300 amplitude are impaired. No single biomarker has been validated yet due to heterogeneity, but continued multimodal research integrating genetics, imaging, and other factors may help establish biomarkers to improve diagnosis and treatment of schizophrenia.
Protocol Presentation of study on Primary CNS lymphomaNarayan Adhikari
Study to evaluate the feasibility of response adapted whole brain radiotherapy after high dose methotrexate based chemotherapy in patients of newly diagnosed primary central nervous system lymphoma
Axonal Polyneuropathy with Anti-Caspr1 Igg1 Paranodal Antibodiessemualkaira
The main electrophysiological manifestation of
autoimmune paranodopathy with anti-contactin-associated protein
1 (Caspr1) positive reported previously was demyelination, with
or without axonal involvement. Here, we reported a patient with
anti-Caspr1 IgG1 positive who had severe axonal involvement accompanied less evidence of demyelination.
This document discusses opportunities and challenges for developing therapies for ALS. It summarizes that over 25 genes have been linked to ALS risk and describes some of the major genes involved like SOD1, TDP43, FUS and C9orf72. Model systems and pathways identified from genes are providing new treatment targets. There are many drugs and approaches in clinical trials. The document outlines the ALS Association's research strategies like establishing a translational program and consortium initiatives to accelerate drug development. Biomarkers, stem cell models and automated microscopy are providing new tools to study mechanisms and do preclinical testing.
The debris of the ‘last major merger’ is dynamically youngSérgio Sacani
The Milky Way’s (MW) inner stellar halo contains an [Fe/H]-rich component with highly eccentric orbits, often referred to as the
‘last major merger.’ Hypotheses for the origin of this component include Gaia-Sausage/Enceladus (GSE), where the progenitor
collided with the MW proto-disc 8–11 Gyr ago, and the Virgo Radial Merger (VRM), where the progenitor collided with the
MW disc within the last 3 Gyr. These two scenarios make different predictions about observable structure in local phase space,
because the morphology of debris depends on how long it has had to phase mix. The recently identified phase-space folds in Gaia
DR3 have positive caustic velocities, making them fundamentally different than the phase-mixed chevrons found in simulations
at late times. Roughly 20 per cent of the stars in the prograde local stellar halo are associated with the observed caustics. Based
on a simple phase-mixing model, the observed number of caustics are consistent with a merger that occurred 1–2 Gyr ago.
We also compare the observed phase-space distribution to FIRE-2 Latte simulations of GSE-like mergers, using a quantitative
measurement of phase mixing (2D causticality). The observed local phase-space distribution best matches the simulated data
1–2 Gyr after collision, and certainly not later than 3 Gyr. This is further evidence that the progenitor of the ‘last major merger’
did not collide with the MW proto-disc at early times, as is thought for the GSE, but instead collided with the MW disc within
the last few Gyr, consistent with the body of work surrounding the VRM.
Authoring a personal GPT for your research and practice: How we created the Q...Leonel Morgado
Thematic analysis in qualitative research is a time-consuming and systematic task, typically done using teams. Team members must ground their activities on common understandings of the major concepts underlying the thematic analysis, and define criteria for its development. However, conceptual misunderstandings, equivocations, and lack of adherence to criteria are challenges to the quality and speed of this process. Given the distributed and uncertain nature of this process, we wondered if the tasks in thematic analysis could be supported by readily available artificial intelligence chatbots. Our early efforts point to potential benefits: not just saving time in the coding process but better adherence to criteria and grounding, by increasing triangulation between humans and artificial intelligence. This tutorial will provide a description and demonstration of the process we followed, as two academic researchers, to develop a custom ChatGPT to assist with qualitative coding in the thematic data analysis process of immersive learning accounts in a survey of the academic literature: QUAL-E Immersive Learning Thematic Analysis Helper. In the hands-on time, participants will try out QUAL-E and develop their ideas for their own qualitative coding ChatGPT. Participants that have the paid ChatGPT Plus subscription can create a draft of their assistants. The organizers will provide course materials and slide deck that participants will be able to utilize to continue development of their custom GPT. The paid subscription to ChatGPT Plus is not required to participate in this workshop, just for trying out personal GPTs during it.
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...Sérgio Sacani
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
(June 12, 2024) Webinar: Development of PET theranostics targeting the molecu...Scintica Instrumentation
Targeting Hsp90 and its pathogen Orthologs with Tethered Inhibitors as a Diagnostic and Therapeutic Strategy for cancer and infectious diseases with Dr. Timothy Haystead.
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...Advanced-Concepts-Team
Presentation in the Science Coffee of the Advanced Concepts Team of the European Space Agency on the 07.06.2024.
Speaker: Diego Blas (IFAE/ICREA)
Title: Gravitational wave detection with orbital motion of Moon and artificial
Abstract:
In this talk I will describe some recent ideas to find gravitational waves from supermassive black holes or of primordial origin by studying their secular effect on the orbital motion of the Moon or satellites that are laser ranged.
Sexuality - Issues, Attitude and Behaviour - Applied Social Psychology - Psyc...PsychoTech Services
A proprietary approach developed by bringing together the best of learning theories from Psychology, design principles from the world of visualization, and pedagogical methods from over a decade of training experience, that enables you to: Learn better, faster!
When I was asked to give a companion lecture in support of ‘The Philosophy of Science’ (https://shorturl.at/4pUXz) I decided not to walk through the detail of the many methodologies in order of use. Instead, I chose to employ a long standing, and ongoing, scientific development as an exemplar. And so, I chose the ever evolving story of Thermodynamics as a scientific investigation at its best.
Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
ESR spectroscopy in liquid food and beverages.pptxPRIYANKA PATEL
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
The technology uses reclaimed CO₂ as the dyeing medium in a closed loop process. When pressurized, CO₂ becomes supercritical (SC-CO₂). In this state CO₂ has a very high solvent power, allowing the dye to dissolve easily.
The binding of cosmological structures by massless topological defectsSérgio Sacani
Assuming spherical symmetry and weak field, it is shown that if one solves the Poisson equation or the Einstein field
equations sourced by a topological defect, i.e. a singularity of a very specific form, the result is a localized gravitational
field capable of driving flat rotation (i.e. Keplerian circular orbits at a constant speed for all radii) of test masses on a thin
spherical shell without any underlying mass. Moreover, a large-scale structure which exploits this solution by assembling
concentrically a number of such topological defects can establish a flat stellar or galactic rotation curve, and can also deflect
light in the same manner as an equipotential (isothermal) sphere. Thus, the need for dark matter or modified gravity theory is
mitigated, at least in part.
1. Thesis Defense for the Degree of
Master of Science in the
Experimental Medicine Program
November 17, 2015
Pierre Zwiegers
Targeted Lentiviral-mediated
Delivery of Progranulin cDNA in a
Genetic Model of Amyotrophic
Lateral Sclerosis
Committee members:
Dr. Christopher A. Shaw
Dr. Shernaz X. Bamji
Dr. Charles Krieger
External Examiner:
Dr. Doris Doudet
Meeting Chair:
Dr. Jason JS Barton
2. Amyotrophic Lateral Sclerosis
Neurotrophic properties of Progranulin and links to ALS
Preliminary data for LV-mediated PGRN upregulation
Research Theme and Objectives
Experimental Methods
Overt Neurobehavioural Measures
End-stage Neuropathology
Decreased mSOD1 Copy Number
Concluding Thoughts & Future Directions
Outline
Background
Results
Summary of Outcomes
Discussion
mSOD1 Copy Number Variation
ALS Models & Pre-clinial Translation
3. Servier medical art modified under a Creative Commons Attribution 3.0 Unported License.
Amyotrophic Lateral Sclerosis
Background
• Age-related and fatally progressive neurodegenerative
condition of upper and lower motor neurons
• World-wide prevalence of 4-7/100 000 population (Chiò
et al. 2013)
• Intractable to therapeutic intervention; Riluzole remains
the only clinically-approved drug (Miller et al. 2012)
Healthy ALS
Upper MNs
Healthy
ALS
Lower MNs
4. Apparently sporadic ALS
90-95%
Familial ALS
5-10%
Turner et al. The Lancet Neurology, Volume 12, Issue 3, 2013, 310 - 322
Amyotrophic Lateral Sclerosis
• More than 180 mutations in the SOD1locus have been linked to ALS (ALSoD, 2015)
• Point mutations primarily result in a toxic gain-of-function property that gives rise
to an ALS phenotype (Harms & Baloh, 2013)
Background
Zwiegers and Shaw. Journal of Con. Biomed Res., Volume 12, Issue 1, 2015, 4-22
• Most widely tested paradigm utilized to model disease and test pre-clinical
efficacy
5. Progranulin and ALS
• PGRN mutations have not been causal to ALS (Petkau & Leavitt, 2014)
• PGRN polymorphisms can result in an earlier onset and a more progressive ALS
phenotype (Sleegers et al, 2008)
Background
▪ Temporal ALS Phenotype Progression
▪ Increased Progranulin Expression
Transgenic
mSOD1 Mouse
Servier medical art modified under a Creative Commons Attribution 3.0 Unported License.
• Early stages of ALS, PGRN levels in the CSF and plasma do not differ from
control patients (Philips et al 2010)
• Post-mortem IHC indicates increased
neuronal and microglial expression
within areas of neurodegeneration
(Irwin et al 2009)
Early stage Late stage
Neuron
Microglia
Progranulin
Philips et al. J Neuropathol and Exp Neurol, Volume 69, Issue 12, 2010, 1191-1200
• PGRN has been shown to be protective in models of Parkinsonism, Alzheimer’s
disease, and arthritis (Minami et al 2014; Van Kampen et al, 2014;Van Kampen
& Kay, 2011, & Tang et al, 2011)
6. Preliminary data for PGRN upregulation
Late-stage targeting of motor neurons in an mSOD1 model preserves neuronal viability
Preliminary Data
Wild-Type G37R
PGRN
WT G37R G37R/PGRN GFP
Background
Provided by Drs. CA Shaw, Denis G. Kay, J Van Kampen & G Lee
7. Hypothesis Research Theme &
Objectives
Degree of ALS phenotype severity
No aberrant
phenotype
Severe
paralysis
Neuron
Microglia
Astrocyte
Early stage PGRN intervention
Can neuronal targeting of exogenous PGRN cDNA at an early stage of the
disease cascade:
Severe
paralysis
lessen the severity of the expressed behavioural phenotype?
attenuate motor neuron loss?
diminish the severity of astrogliosis and microgliosis?
8. Lentiviral –mediated transgene delivery
Servier medical art modified under a Creative Commons Attribution 3.0 Unported License.
Experimental Methods
G37R (line 29)/ WT mice
at 3.5-4 months of age
Transgene integration;
under control of CMV
promoter
Histological Assays
• Microgliosis (Iba-1)
• Astrogliosis (GFAP)
• Neuronal Viability &
Morphology (Cresyl Violet, ChAT)
Monitoring/Neurobehavioural Assays
• Leg Extension Reflex Test
• Latency to fall from an
elevated grid
• Weight
5x2µL injections @
4.0x108 TU/mL VSV-G
pseudotyped
lentiviral vector
encoding
GFP/PGRN into
gastrocnemius
muscles
1 week 2 weeks 4 weeks
9. Overt Neurobehavioural Measures
Results
Weight
• Male G37R animals presented with a delay in phenotype-related weight loss
• PGRN administration did not attenuate the progressive loss of body mass
.. observed in Tg mSOD1 animals
Leg Extension & Wire Hang Test
• Early-stage PGRN delivery did not ameliorate outcomes in either measure
• Male animals showcased a more severe deficit at earlier time points e ap
10. Overt Neurobehavioural Measures
Results
Onset & Survival
• PGRN cDNA delivery did not delay disease onset or prolong survival
• Male Tg animals showed evidence of an attenuated disease progression
Median Onset (d)
GFP: 344
PGRN: 315
Median Survival (d)
GFP: 609
PGRN: 607
Median Onset (d)
GFP: 287
PGRN: 294
Median Survival (d)
GFP: 582
PGRN: 573
Male
Female
11. End-stage Neuropathology
Results
Exogenous transgene cDNA
• no qPCR signal for PGRN/GFP in formalin-fixed L3-L5 tissue specimens
• no immunohistological evidence for GFP expression in region L3-L5
Neuronal Morphology
• Tg male animals presented with neurons of a smaller diameter
• Early-stage PGRN intervention had no effect on neuronal morphology
† Healthy
ǂ Atrophying
Atrophying neurons
Cumulative
%Distribution
Healthy neurons
Diameter (µm)
ǂ
ǂ
ǂ
ǂ
ǂ
†
†
Diameter (µm)
12. End-stage Neuropathology
Results
Exogenous transgene cDNA
• no qPCR signal for PGRN/GFP in formalin-fixed L3-L5 tissue specimens
• no immunohistological evidence for GFP expression in region L3-L5
Neuronal Morphology
• Tg male animals presented with neurons of a smaller diameter
• Early-stage PGRN intervention had no effect on neuronal morphology
Neuronal Viability
• Male mSOD1 animals showed reduced neuronal viability
• PGRN administration did not diminish neuronal loss in the targeted L3-L5 region
αnti-Choline acetyltransferaseCresyl Violet Assay
Lumbar Spinal Cord
Level
13. End-stage Neuropathology
Results
Neuroinflammation
• Tg mSOD1 animals showed a significant increase in neuroinflammatory
processess
• Early-stage PGRN cDNA delivery did not attenuate microglial/astrocytic
activation
Astrogliosis Microgliosis
14. LV-mediated Transgene Delivery
No aberrant
phenotype
Severe
paralysis
Early stage PGRN intervention Late stage PGRN intervention
- No effect on disease
onset or duration
- No mitigation of neuronal
loss/neuroinflammation
- Preserved ChAT positive
motor neurons
Summary of Outcomes
How do we reconcile these paradoxical outcomes?
Δ time
• Transgene presence not confirmed at end point
- early targeting may have altered expression levels within neuronal
populations more susceptible to the neurotoxic insult
15. LV-mediated Transgene Delivery
Summary of Outcomes
How do we reconcile these paradoxical outcomes?
• Transgene presence not confirmed at end point
- early targeting may have altered expression levels within neuronal
populations more susceptible to the neurotoxic insult
• Use of deficient mSOD1 model animals
Disease Severity
No aberrant
phenotype
Severe
paralysis
Early stage PGRN intervention Late stage PGRN intervention
- No effect on disease
onset or duration
- No mitigation of neuronal
loss/neuroinflammation
- Preserved ChAT positive
motor neurons
17. Changes in mSOD1copy number
Results
Age (d)
• G37R animals generated for this
study showed a 46% lifespan increase
• Intra-sex comparisons showed a
significant colony effect (p<0.0001)
• Intra-colony comparisons indicated a
significant difference in the relative
abundance of the mutant locus
• Commercially-derived animals
showed a more uniform distribution in
ΔCT values; with a near 3-fold reduc-
tion in mSOD1 copy number
*** ***
***** Near 3-fold
reduction
Colony
Source: Collaborator Commercial
18. Murine SOD1 ALS Model Systems
Fertilized Egg
Integrated
mutant human
SOD1 locus
Pseudopregnant
mouse
Established transgenic lines
▪ Degree of transgene integration
▪ Phenotypic disease severity
Rare recombination events
Varied disease presentation
Transgene level variation
Discussion
Zwiegers et al. J. of Negative Results in Biomedicne, Volume 13, Issue 1, 2014
Servier medical art modified under a Creative Commons Attribution 3.0 Unported License.
19. Discussion
Original Founder Line
Line Deposited at Repository
Animals Generated for Breeding
Progeny Generated for Study
Deposited with an uncharacterized
drop in copy number
Changes in mSOD1copy number
Data provided by the Jackson Laboratory
Servier medical art modified under a Creative Commons Attribution 3.0 Unported License.
Historical Controls
20. • With > 50 RCTs, positive pre-clinical findings have not been recapitulated in the
clinic (Mitsumoto et al. 2014)
Pre-clinical
mSOD1 Model
Patient Population
Putative
therapeutics
Pre-clinical Translational Studies
Discussion
21. • With > 50 RCTs, positive pre-clinical findings have not been recapitulated in the
clinic (Mitsumoto et al. 2014)
Pre-clinical
mSOD1 Model
Patient Population
Putative
therapeutics
• mSOD1models do not serve as a predictor of clinical success
Pre-clinical Translational Studies
Discussion
22. - models a small % of ALS cases; logistical considerations for clinical studies
Why are mSOD1models a poor predictor of clinical success?
Zwiegers and Shaw. Journal of Con. Biomed Res., Volume 12, Issue 1, 2015, 4-22
Pre-clinical Translational Studies
Discussion
23. - models a small % of ALS cases; logistical considerations for clinical studies
- lack of replicative studies prior to clinical translation
Why are mSOD1models a poor predictor of clinical success?
Turner et al. The Lancet Neurology, Volume 12, Issue 3, 2013, 310 - 322
Pre-clinical Translational Studies
Discussion
24. - models a small % of ALS cases; logistical considerations for clinical studies
- lack of replicative studies prior to clinical translation
- pre-clinical outcomes may just be a measurement of experimental noise
(Scott et al, 2008)
Why are mSOD1models a poor predictor of clinical success?
Turner et al. The Lancet Neurology, Volume 12, Issue 3, 2013, 310 - 322
Pre-clinical Translational Studies
Discussion
Zwiegers et al. J. of Negative Results in Biomedicne, Volume 13, Issue 1, 2014
mSOD1 Copy Number
Lifespan
Phenotype
Severity
Increasing transgene dependent severity of ALS
SOD1 low-copy number SOD1 high-copy number
↓ likelihood of (+)ve outcome↑ likelihood of (+)ve outcome
25. Conclusion
Servier medical art modified under a Creative Commons Attribution 3.0 Unported License.
• Must adopt a multi-modal approach to test pre-clinical efficacy prior to
translational efforts
Putative therapeutics
ALS Patient
PopulationPre-clinical ALS Models
Invertebrate models Patient-derived Nerve Cells
• Critical need to publish negative data so that non-productive research efforts
can be identified/addressed
• Pre-clinical ALS studies need to publish findings in the context of loci copy
number
• Research community needs to incentivize independent replicative studies prior
to any clinical translation
26. References
ALSoD, 2015. ALS Online Genetics Database. [online] Available at: http://alsod.iop.kcl.ac.uk/Index.aspx. Available at: http://alsod.iop.kcl.ac.uk/.
Chiò, a. et al., 2013. Global epidemiology of amyotrophic lateral sclerosis: A systematic review of the published literature. Neuroepidemiology, 41(2), pp.118–130.
Harms, M.B. & Baloh, R.H., 2013. Clinical Neurogenetics: Amyotrophic Lateral Sclerosis. Neurologic Clinics, 31(4), pp.929–950
Irwin, D., Lippa, C.F. & Rosso, a., 2009. Progranulin (PGRN) expression in ALS: An immunohistochemical study. Journal of the Neurological Sciences, 276(1-2), pp.9–13. Available
at: http://dx.doi.org/10.1016/j.jns.2008.08.024.
Miller RG, JD M, Moore DH. Riluzole for amyotrophic lateral sclerosis (ALS)/motor neuron disease (MND). Cochrane Database of Syst Rev 2012.
Mitsumoto, H., Brooks, B.R. & Silani, V., 2014. Clinical trials in amyotrophic lateral sclerosis: why so many negative trials and how can trials be improved? The Lancet Neurology,
13(11), pp.1127–1138. Available at: http://linkinghub.elsevier.com/retrieve/pii/S1474442214701292.
Petkau, T.L. & Leavitt, B.R., 2014. Progranulin in neurodegenerative disease. Trends in Neurosciences, 37(7), pp.388–398. Available at:
http://dx.doi.org/10.1016/j.tins.2014.04.003.
Philips, T. et al., 2010. Microglial upregulation of progranulin as a marker of motor neuron degeneration. Journal of neuropathology and experimental neurology, 69(12),
pp.1191–1200.
Ryan, C.L. et al., 2009. Progranulin is expressed within motor neurons and promotes neuronal cell survival. BMC neuroscience, 10, p.130.
Scott, S. et al., 2008. Design, power, and interpretation of studies in the standard murine model of ALS. Amyotrophic lateral sclerosis : official publication of the World Federation
of Neurology Research Group on Motor Neuron Diseases, 9(1), pp.4–15.
Sleegers, K. et al., 2008. Progranulin genetic variability contributes to amyotrophic lateral sclerosis. Neurology, 71, pp.253–259.
Tang, W. et al., 2011. The growth factor progranulin binds to TNF receptors and is therapeutic against inflammatory arthritis in mice. Science (New York, N.Y.), 332(6028), pp.478–
484
Turner, M.R. et al., 2013. Mechanisms, models and biomarkers in amyotrophic lateral sclerosis. Amyotrophic lateral sclerosis & frontotemporal degeneration, 14 Suppl 1, pp.19–
32. Available at: http://www.ncbi.nlm.nih.gov/pubmed/23678877.
Van Kampen, J. & Kay, D., 2011. Progranulin gene therapy prevents plaque formation and synapse loss in a rodent model of AD. Alzheimer’s & Dementia, 7(4), pp.e7–e8.
Available at: http://dx.doi.org/10.1016/j.jalz.2011.09.021.
Van Kampen, J.M., Baranowski, D. & Kay, D.G., 2014. Progranulin gene delivery protects dopaminergic neurons in a mouse model of Parkinson’s disease. PLoS ONE, 9(5).
Zwiegers, P., Lee, G. & Shaw, C. a, 2014. Reduction in hSOD1 copy number significantly impacts ALS phenotype presentation in G37R (line 29) mice: implications for the
assessment of putative therapeutic agents. Journal of Negative Results in BioMedicine, 13(1), p.14. Available at: http://www.jnrbm.com/content/13/1/14.
Zwiegers, P. & Shaw, C.A., 2015. Disparity of outcomes : the limits of modeling amyotrophic lateral sclerosis in murine models and translating results clinically. Journal of
Controversies in Biomedical Research, 1(1), pp.4–22.
29. Methods
• 10μm-thick, 100μm apart tissue sections were compared to a spinal cord atlas and organized
accordingly
Spinal Cord Organization
L1 L2 L3 L4 L5 L6
Ventral Horn
Cuneate & Gracile
fasiculus
Dorsal Horn
30. Methods
Neuronal Viability
Small motor neurons:
Area = 130-240um2
VH
CC
Large motor neurons:
Area = 240-950 um2
• The VH of cresyl violet stained tissues sections were captured at 10x
• Colour inverted, contrast between objects enhanced
• Image analysed by CellProfiler software; based on size criteria
• Experimenter manually confirmed that identified objects were not artefacts
Add details re:comparison of MN sizes; Friese et al paper incl
31. Methods
Neuronal Morphology
• Targeted the VH of cresyl violet stained tissues sections captured at 40x
• Cell diameter estimated by bisecting cell of interest by two lines; size measured
with ImageJ
• “Healthy” neurons:
- prominent nucleolus; definitive neuron-like morphology
- 1-5 processes present within the plane of sectioning
• “Atrophying” neurons:
- neuronal shrinkage
- hyperchromatic appearance
ǂ
ǂ
ǂ
ǂ
†
†
ǂ Atrophying MN
† Apparently Healthy MN
37. Supplementary Slides
Friese, A. et al. Gamma and alpha motor neurons distinguished by expression of transcription factor Err3.
Proceedings of the National Academy of Sciences 106, 13588–13593 (2009).
39. Supplementary Slides
LV-mediated cDNA Expression
A
20x
D
20x
B
40x
E
40x
F
100x
C
100x
100x
LV-PGRN
100x
LV-SCRB
20x
LV-SCRB
20x
LV-PGRN
Produced by MS Petrik & R Cruz-Aguado
GFP and ChAT immunolabeling following
gastroc-mediated LV injection show viral
delivery to MN
LV-SCRB
LV-PGRN
40. Supplementary Slides
• No significant difference in mSOD1 copy number between treatment groups
mSOD1 Transgene Presence
42. Jan 2012
Grad program start
Dec 2010- Mar 2011
Establish a breeding colony
& generate a cohort of
experimental animals.
May 2011
Weekly baseline assessment at 2-2.5 months of age
Jun + Aug 2011
Bi-lateral LV injections at
3-3.5 months of age
Jun 2011 – Jan 2013
Continue acquisition of behavioral data:
Late 2012 – Mar 2013
euthanize animals exhibiting
signs of severe ALS
Servier medical art modified under a Creative Commons Attribution 3.0 Unported License.
Behavioural Measures
Histological Processing &
Measurements
EXPERIMENTAL TIMELINE
Aug 2013 – Apr 2015
Tissue processing and post-study histological
assays.
Antal et al 2007
• Injection Sites
Weight
Leg Extension Reflex Score
Wire Hang
Neuronal Viability & Morphology
Astrogliosis
Microgliosis
Experimental Timeline
43. Results
Male
Female
• PGRN administration in Tg mSOD1 animals did not attenuate the disease-related weight loss
• Male Tg animals appeared to exhibit a delayed phenotypic progression compared to females
9 10 11 12 13 18 18 19 21 23 53 57 61 64 65 67 69 71 73 75 78 79 81 83 86 87 89
9 10 11 12 13 18 18 19 21 23 53 57 61 64 65 67 69 71 73 75 78 79 81 83 86 87 89
Average Age (weeks)
Average Age (weeks)
Weight (Mean +/- SEM)
44. • PGRN administration did not mediate a positive effect in the LE reflex score
• Tg mSOD1 animals started to show deficits at later time points
9 10 11 12 13 18 18 19 21 23 57 61 64 65 67 69 71 73 75 78 79 81 83 86 87 89
Average Age (weeks)
25 27 29 31 33 35 37 40 41 43 45 47 49 51 53
Average Age (weeks)
Male
Female
ResultsLeg Extension reflex (Mean +/- SEM)
9 10 11 12 13 18 18 19 21 23 57 61 64 65 67 69 71 73 75 78 79 81 83 86 87 8925 27 29 31 33 35 37 40 41 43 45 47 49 51 53
45. • Male Tg animals appeared to exhibit an earlier deficit in comparison to female counterparts
• PGRN administration in Tg mSOD1 animals mediated no positive effect on the measure
Results
9 10 11 12 13 18 18 19 21 23 57 61 64 65 67 69 71 73 75 78 79 81 83 86 87 89
Average Age (weeks)
25 27 29 31 33 35 37 40 41 43 45 47 49 51 53
9 10 11 12 13 18 18 19 21 23 57 61 64 65 67 69 71 73 75 78 79 81 83 86 87 89
Average Age (weeks)
25 27 29 31 33 35 37 40 41 43 45 47 49 51 53
Male
Female
Wire Hang Test (Mean +/- SEM)
46. Neuronal Viability & Morphology Results
Atrophying neurons
Cumulative
%Distribution
Healthy neurons
Diameter (µm)
• In Tg mSOD1 animals, PGRN administration did not affect the size distribution of motor
neuron cell bodies
Mean Neuronal Size (Mean +/- SD)
47. Neuronal Viability & Morphology (Mean +/- SD) Results
Small motor neurons:
Area = 130-240um2
VH
CC
Large motor neurons:
Area = 240-950 um2
• Early-stage PGRN administration mediated no significant effect on end-stage neuronal viability
throughout the lumbar spinal cord
48. Neuronal Viability & Neuroinflammation (Mean +/- SD) Results
• Tg male mSOD1 animals showed significantly reduced MN numbers in the targeted L3-L5
region as well as an increase in neuroinflammatory cells
• PGRN administration at an early stage did not ameliorate MN loss or gliosis
49. Mean signal 33.47 34.01 12.6 12.27
SD 11.55 10.91 2.535 6.122
n 12 6 24 23
• Tg animals that were generated from Jax experienced a drop in copy number prior to being
deposited at the repository
• Decreased copy number resulted in an attenuated disease presentation, with the end point
more than 15 months following lentiviral administration, compared to a colony generated
from a local collaborator
Editor's Notes
Significant motor neuron loss ultimately leads to muscle atrophy, with death due to respiratory complications 2-4 years following disease onset (Chiò et al. 2013)
Clinically, Riluzole only presents with a modest 2-3 month increase in overall lifespan
• L.O.F and the resulting haploinsufficiency implicated in tau negative, ubiquitin
positive FTLD (Cruts et al., 2006; Baker et al. 2006)
Introduce with the fact that we had pre-lim in vitro evidence suggesting a positive effect
Mention which regions of the LSc you are focusing on for your counts
Explain hoe you were able to do that
5x2µL injections @ 4.0x108 TU/mL
VSV-G pseudotyped lentiviral vector encoding GFP/PGRN into gastrocnemius muscles
Weight:
• Earliest parameter distinguishing between WT and Tg cohorts
Weight:
• Earliest parameter distinguishing between WT and Tg cohorts
Tg animals 46% inc in lifespan compared to prototypical model
When comparing original breeders, turns out they are the same as historical controls therefore drop happened when line deposited at repository
Tg animals 46% inc in lifespan compared to prototypical model
When comparing original breeders, turns out they are the same as historical controls therefore drop happened when line deposited at repository
Transgenic anmal models overexressing mutant SOD1 have become the most widely utilized pre-clinical model to test ALS therapeutics
However, these have various issues that inhihibit clinical applications
Transgene variation
Not modeling a wide swath of pts
Not a good predictor of clinical efficacy
Significant overexpression of mutant locus (i.e. non-physiologically relevant levels)
Might want to have a slide ready that details the CTs for the progeny from each male breeder; also have explanatory slides ready for the IHC quantification
What were the housekeeping genes for these assays???
Also, what about including the dCT values you have for the few breeders that you did do???
The antithesis also holds: when parametric issues are resolved, the lack of a preclinical effect does model the clinical outcome. This finding is likely consequent to severe transgene overexpression and short of inhibiting SOD1 expression/misfolding, any treatment will fail to be efficacious
Mean +/- SEMALSoD, 2015. ALS Online Genetics Database. [online] Available at: http://alsod.iop.kcl.ac.uk/Index.aspx. Available at: http://alsod.iop.kcl.ac.uk/.
Chiò, a. et al., 2013. Global epidemiology of amyotrophic lateral sclerosis: A systematic review of the published literature. Neuroepidemiology, 41(2), pp.118–130.
Harms, M.B. & Baloh, R.H., 2013. Clinical Neurogenetics: Amyotrophic Lateral Sclerosis. Neurologic ……..Clinics, 31(4), pp.929–950
Irwin, D., Lippa, C.F. & Rosso, a., 2009. Progranulin (PGRN) expression in ALS: An immunohistochemical study. Journal of the Neurological Sciences, 276(1-2), pp.9–13. Available at: http://dx.doi.org/10.1016/j.jns.2008.08.024.
Miller RG, JD M, Moore DH. Riluzole for amyotrophic lateral sclerosis (ALS)/motor neuron disease ……..(MND). Cochrane Database of Syst Rev 2012; CD001447.
Mitsumoto, H., Brooks, B.R. & Silani, V., 2014. Clinical trials in amyotrophic lateral sclerosis: why so many negative trials and how can trials be improved? The Lancet Neurology, 13(11), pp.1127–1138. Available at: http://linkinghub.elsevier.com/retrieve/pii/S1474442214701292.
Petkau, T.L. & Leavitt, B.R., 2014. Progranulin in neurodegenerative disease. Trends in Neurosciences, 37(7), pp.388–398. Available at: http://dx.doi.org/10.1016/j.tins.2014.04.003.
Philips, T. et al., 2010. Microglial upregulation of progranulin as a marker of motor neuron degeneration. Journal of neuropathology and experimental neurology, 69(12), pp.1191–1200.
Ryan, C.L. et al., 2009. Progranulin is expressed within motor neurons and promotes neuronal cell survival. BMC neuroscience, 10, p.130.
Scott, S. et al., 2008. Design, power, and interpretation of studies in the standard murine model of ALS. Amyotrophic lateral sclerosis : official publication of the World Federation of Neurology Research Group on Motor Neuron Diseases, 9(1), pp.4–15.
Sleegers, K. et al., 2008. Progranulin genetic variability contributes to amyotrophic lateral sclerosis. Neurology, 71, pp.253–259.
Tang, W. et al., 2011. The growth factor progranulin binds to TNF receptors and is therapeutic against inflammatory arthritis in mice. Science (New York, N.Y.), 332(6028), pp.478–484
Turner, M.R. et al., 2013. Mechanisms, models and biomarkers in amyotrophic lateral sclerosis. Amyotrophic lateral sclerosis & frontotemporal degeneration, 14 Suppl 1, pp.19–32. Available at: http://www.ncbi.nlm.nih.gov/pubmed/23678877.
Van Kampen, J. & Kay, D., 2011. Progranulin gene therapy prevents plaque formation and synapse loss in a rodent model of AD. Alzheimer’s & Dementia, 7(4), pp.e7–e8. Available at: http://dx.doi.org/10.1016/j.jalz.2011.09.021.
Van Kampen, J.M., Baranowski, D. & Kay, D.G., 2014. Progranulin gene delivery protects dopaminergic neurons in a mouse model of Parkinson’s disease. PLoS ONE, 9(5).
Zwiegers, P., Lee, G. & Shaw, C. a, 2014. Reduction in hSOD1 copy number significantly impacts ALS phenotype presentation in G37R (line 29) mice: implications for the assessment of putative therapeutic agents. Journal of Negative Results in BioMedicine, 13(1), p.14. Available at: http://www.jnrbm.com/content/13/1/14.
Zwiegers, P. & Shaw, C.A., 2015. Disparity of outcomes : the limits of modeling amyotrophic lateral sclerosis in murine models and translating results clinically. Journal of Controversies in Biomedical Research, 1(1), pp.4–22.
included the size and shape of the ventral horn, gracile fasiculus, and spinal lamina
Large motor neurons: equivalent diameter = 8-16pixels
Small motor neurons: equivalent diameter = 6-8pixels
Large motor neurons: equivalent diameter = 8-16pixels
Small motor neurons: equivalent diameter = 6-8pixels
Large motor neurons: equivalent diameter = 8-16pixels
Small motor neurons: equivalent diameter = 6-8pixels
Large motor neurons: equivalent diameter = 8-16pixels
Small motor neurons: equivalent diameter = 6-8pixels
Large motor neurons: equivalent diameter = 8-16pixels
Small motor neurons: equivalent diameter = 6-8pixels
Mean cross sectional area
Red fluorescence is ChAT and yellowish green is the virus. The virus is present in the majority of motor neurons in both sets of injected animals, and is present in BOTH ventral horns. Arrows indicate motor neurons that do not appear to have the virus present. Under 100x magnification, LV-SCRB (scrambled) injected animals exhibit robust and triangular motor neuron structure and localized virus labeling; whereas, notably in LV-PGRN injected animals, GFP and ChAT labeling appeared diffuse, scattered and lacking a distinct nucleus in some motor neurons. Cell counts did not show any significance, however, it must be emphasized that n=2 per group. All these animals were fed BSSG.
Red fluorescence is ChAT and yellowish green is the virus. The virus is present in the majority of motor neurons in both sets of injected animals, and is present in BOTH ventral horns. Arrows indicate motor neurons that do not appear to have the virus present. Under 100x magnification, LV-SCRB (scrambled) injected animals exhibit robust and triangular motor neuron structure and localized virus labeling; whereas, notably in LV-PGRN injected animals, GFP and ChAT labeling appeared diffuse, scattered and lacking a distinct nucleus in some motor neurons. Cell counts did not show any significance, however, it must be emphasized that n=2 per group. All these animals were fed BSSG.