American Society of Hematology 75th Annual Conference. Accelerated Repair of Vascular Injury in Diabetes by TGF-B1 Modified Hematopoietic Stem Cells (HSC)
Eduardo Marbán, MD, PhD gives a deep dive into the complexities of regenerative cardiovascular medicine and the future directions for cell therapies.
Dr. Marbán’s lab has found several noncoding RNA (ncRNA) species, including short Y RNAs, which themselves have intriguing biological actions. The ncRNA within extracellular vesicles can either be used as they occur naturally or serve as bioinspiration for new chemical entities. The next generations of cell-free biologics (extracellular vesicles and noncoding RNAs) may provide, or even transcend, the benefits of cell therapy without the intrinsic limitations.
Key Topics Include:
- Understand the current status of cell therapy for heart disease
- Review the mechanisms whereby cells exert their therapeutic benefits
- Explore the future of RNA drugs inspired by the contents of extracellular vesicles
Summary of ADC Targets For Solid Tumors & Hematological Tumors.pdfDoriaFang
Currently, popular targets include CD family, BCMA, HER2, TROP2, Tissue factor, Nectin-4, FRα, EGFR, etc. Here, we briefly introduce ADC targets in solid tumors and hematological tumors.
Eduardo Marbán, MD, PhD gives a deep dive into the complexities of regenerative cardiovascular medicine and the future directions for cell therapies.
Dr. Marbán’s lab has found several noncoding RNA (ncRNA) species, including short Y RNAs, which themselves have intriguing biological actions. The ncRNA within extracellular vesicles can either be used as they occur naturally or serve as bioinspiration for new chemical entities. The next generations of cell-free biologics (extracellular vesicles and noncoding RNAs) may provide, or even transcend, the benefits of cell therapy without the intrinsic limitations.
Key Topics Include:
- Understand the current status of cell therapy for heart disease
- Review the mechanisms whereby cells exert their therapeutic benefits
- Explore the future of RNA drugs inspired by the contents of extracellular vesicles
Summary of ADC Targets For Solid Tumors & Hematological Tumors.pdfDoriaFang
Currently, popular targets include CD family, BCMA, HER2, TROP2, Tissue factor, Nectin-4, FRα, EGFR, etc. Here, we briefly introduce ADC targets in solid tumors and hematological tumors.
Use of TEG® in Acute Traumatic Brain Injurykathleenmccann
Acute traumatic coagulopathy complicate 25 – 75% of cases of traumatic brain injury, incidence increasing with severity of injury. The complex processes that cause this are a culmination of the activation of anticoagulant and fibrinolytic pathways. Much has been published about the clinical outcomes of brain injury and the associated coagulopathy with platelet dysfunction. The mechanism, however, is still not understood.
Clinical management of these patients to date relies on traditional serum based screens: PT, INR, PTT, platelet counts, fibrinogen, and D-dimer assays. These values do not reflect whole blood hemostasis nor do they reflect a cell based model of hemostasis. Thromboelastography with platelet mapping, however, provides real-time data on:
Coagulation pathway function
Fibrinogen function
Platelet function
Fibrinolysis
We purport the expanded use of TEG® as a viscoelastic point-of-care test of whole blood which more accurately reflects hemostasis. It determines the hemostatic integrity of whole blood in vitro by giving real-time information on thrombus initiation, amplification, propagation, and termination. It has long been used in management in transplant patients and during cardiac surgery. It has recently become increasingly used in the management of trauma patients, including those presenting with traumatic brain injury.
We show that TEG® is useful not only in guiding treatment, but also in studying the mechanisms underlying acute traumatic coagulopathy and platelet dysfunction in cases of hemorrhagic brain injury, in both the animal model and in the clinical setting.
Effect of stemregenin1 and sb431542 small molecules on ex vivo expansion of u...Liberty University (LU)
Cord blood Hematopoietic stem cells (HSCs) with several advantages including low chance of viral contamination and low rate of Graft versus host disease (GVHD) are appropriate candidate for vast medical applications such as transplantation. The main obstacle of cord blood HSCs is the low number cells. To improve ex-vivo expansion of umbilical cord HSCs we introduced a new culture system. Isolated HSCs were seeded in Three-dimensional(3D) on Polyethersulfone(PES) scaffolds and Twodimensional(2D) culture conditions and treated with SB431542 and Stemregenin1(SR1) small molecules. On the fifth and tenth days the expanded cells in different groups were investigated for expression of specific markers by flow cytometry, expression of some stemness genes by qRT-PCR and colony formation by methocult medium. SR1 molecule significantly increased expansion of CD34+ cells while SB431542 induced more CD34+/38+ cells. Also SB431542 treated cells showed higher colony formation capacity. SR1 increased the expression of c-Myc, HOXB4 and SALL4 while SB431542 seemed to inhibit HOXB4 expression and increase SALL4.Together all, this study introduced a new ex vivo culture setting for further medical application of HSCs. Our data showed simultaneous use of these two small molecules can provide appropriate outcome for HSCs transplantation includes both of engraftment and repopulation.
The article deals with the molecular and insilico analysis of compound heterozygosity to understand the complex interactions and effects of the genetic compounds in HCM.
Antibody drug conjugates (ADCs) are undoubtedly one of the hottest research areas in recent years. To date, there are 15 approved ADCs in the world, of which 13 have been approved by the FDA, one in China (Disitamab Vedotin) and one in Japan (Akalux).
Bio-Markers of Heart Failure (Dr.LIKHIT T)Likhit T
A brief on bio-markers of Heart failure...First of all, I thank the Authors of all the books from which I picked the points to make this presentation.. This presentation includes classification of bio-markers and explanation according their importance.. Thank you
BCMACD3 Bispecific Antibodies for Multiple Myeloma (MM).pdfDoriaFang
Currently, BCMA/CD3 bispecific antibodies have shown therapeutic potential in R/R MM. Here, we will explore the BCMA/CD3 bispecific antibodies under investigation for treating MM.
Summary of PROTAC And Other Targeted Protein Degradation Technologies.pdfDoriaFang
We summarize various targeted degradation strategies and their respective advantages and disadvantages, hoping to provide guidance value for the development of targeted protein degradation drugs.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Use of TEG® in Acute Traumatic Brain Injurykathleenmccann
Acute traumatic coagulopathy complicate 25 – 75% of cases of traumatic brain injury, incidence increasing with severity of injury. The complex processes that cause this are a culmination of the activation of anticoagulant and fibrinolytic pathways. Much has been published about the clinical outcomes of brain injury and the associated coagulopathy with platelet dysfunction. The mechanism, however, is still not understood.
Clinical management of these patients to date relies on traditional serum based screens: PT, INR, PTT, platelet counts, fibrinogen, and D-dimer assays. These values do not reflect whole blood hemostasis nor do they reflect a cell based model of hemostasis. Thromboelastography with platelet mapping, however, provides real-time data on:
Coagulation pathway function
Fibrinogen function
Platelet function
Fibrinolysis
We purport the expanded use of TEG® as a viscoelastic point-of-care test of whole blood which more accurately reflects hemostasis. It determines the hemostatic integrity of whole blood in vitro by giving real-time information on thrombus initiation, amplification, propagation, and termination. It has long been used in management in transplant patients and during cardiac surgery. It has recently become increasingly used in the management of trauma patients, including those presenting with traumatic brain injury.
We show that TEG® is useful not only in guiding treatment, but also in studying the mechanisms underlying acute traumatic coagulopathy and platelet dysfunction in cases of hemorrhagic brain injury, in both the animal model and in the clinical setting.
Effect of stemregenin1 and sb431542 small molecules on ex vivo expansion of u...Liberty University (LU)
Cord blood Hematopoietic stem cells (HSCs) with several advantages including low chance of viral contamination and low rate of Graft versus host disease (GVHD) are appropriate candidate for vast medical applications such as transplantation. The main obstacle of cord blood HSCs is the low number cells. To improve ex-vivo expansion of umbilical cord HSCs we introduced a new culture system. Isolated HSCs were seeded in Three-dimensional(3D) on Polyethersulfone(PES) scaffolds and Twodimensional(2D) culture conditions and treated with SB431542 and Stemregenin1(SR1) small molecules. On the fifth and tenth days the expanded cells in different groups were investigated for expression of specific markers by flow cytometry, expression of some stemness genes by qRT-PCR and colony formation by methocult medium. SR1 molecule significantly increased expansion of CD34+ cells while SB431542 induced more CD34+/38+ cells. Also SB431542 treated cells showed higher colony formation capacity. SR1 increased the expression of c-Myc, HOXB4 and SALL4 while SB431542 seemed to inhibit HOXB4 expression and increase SALL4.Together all, this study introduced a new ex vivo culture setting for further medical application of HSCs. Our data showed simultaneous use of these two small molecules can provide appropriate outcome for HSCs transplantation includes both of engraftment and repopulation.
The article deals with the molecular and insilico analysis of compound heterozygosity to understand the complex interactions and effects of the genetic compounds in HCM.
Antibody drug conjugates (ADCs) are undoubtedly one of the hottest research areas in recent years. To date, there are 15 approved ADCs in the world, of which 13 have been approved by the FDA, one in China (Disitamab Vedotin) and one in Japan (Akalux).
Bio-Markers of Heart Failure (Dr.LIKHIT T)Likhit T
A brief on bio-markers of Heart failure...First of all, I thank the Authors of all the books from which I picked the points to make this presentation.. This presentation includes classification of bio-markers and explanation according their importance.. Thank you
BCMACD3 Bispecific Antibodies for Multiple Myeloma (MM).pdfDoriaFang
Currently, BCMA/CD3 bispecific antibodies have shown therapeutic potential in R/R MM. Here, we will explore the BCMA/CD3 bispecific antibodies under investigation for treating MM.
Summary of PROTAC And Other Targeted Protein Degradation Technologies.pdfDoriaFang
We summarize various targeted degradation strategies and their respective advantages and disadvantages, hoping to provide guidance value for the development of targeted protein degradation drugs.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
Basavarajeeyam is an important text for ayurvedic physician belonging to andhra pradehs. It is a popular compendium in various parts of our country as well as in andhra pradesh. The content of the text was presented in sanskrit and telugu language (Bilingual). One of the most famous book in ayurvedic pharmaceutics and therapeutics. This book contains 25 chapters called as prakaranas. Many rasaoushadis were explained, pioneer of dhatu druti, nadi pareeksha, mutra pareeksha etc. Belongs to the period of 15-16 century. New diseases like upadamsha, phiranga rogas are explained.
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Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
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ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
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1. Accelerated Repair of Vascular Injury in Diabetes by TGF-β1 Modified Hematopoietic Stem Cells (HSC)
Stephen Bartelmez1, Francis Ruscetti 2, Patrick Iversen3, Gail Gannon1, Aqeela Afzal4, Ashay Bhatwadekar4, Maria Grant4
1BetaStem Therapeutics, San Francisco CA, 2Lab. of Leukocyte Biology, NCI, Frederick, MD, 3AVI-Biopharma, Corvallis, OR, 4University of Florida, Gainesville, FL
Abstract No. 1398
INTRODUCTION METHODS CONCLUSIONS
Isolation of human CD34+ cells: Blood mononuclear cells (MNC) were enriched by F/H separation. 3.3 X 107 MNCs and blocked with FcR blocking reagent
Diabetic retinopathy induces retinal vascular endothelial damage resulting in ischemia, Figure 6: Vascular Repair
1) Our initial studies indicate that blood CD34+
(Miltenyi Biotec Inc., Auburn, CA) , and 33 µl of magnetic microbeads conjugated with an anti-CD34 antibody was added. CD34+ cells were positively selected using
edema, hemorrhage and eventually blindness. We have recently shown that CD34+ /EPC is increased by Blocking
cells (and plasma) from diabetic patients
an automated magnetic selection autoMACS™ (Miltenyi Biotec Inc.). CD34+ purity was determined by co-staining with PE-anti-CD34 (high) (Miltenyi Biotec Inc.)
from diabetic patients are defective in their ability to repair damaged vessels and their TGF-β in both Diabetic and
and FITC-anti-CD45 (mid). have increased levels of TGF-β. Non-diabetic CD34+ cells
migratory response to SDF-1 is markedly reduced. We describe a new approach to treat TGF-beta specific (qRT-PCR) Analyses of peripheral blood CD34+ cells: Total RNA was isolated using TRI reagent (Sigma-Aldrich, USA). The RNA samples were 2) Transient blockade of TGF-β in CD34+ cells (from Fig 2)
retinopathy in which the patient’s hematopoietic stem cells (HSC) are greatly enhanced to converted to single strand cDNA using the high capacity DNA archive kit (Applied Biosystems, Foster City, CA). Taqman universal master mix (Applied Biosystems)
by PMO enhances vasculature reparative
was added to cDNA for real-time PCR analyses.
home to damaged retinal vessels and generate endothelial progenitor cells (EPCs) which
function by diabetic and nondiabetic CD34+
TGF-b blockade: CD34+ cells were incubated for 8 hrs at 37°C with either 40 ug/ml TGF-β PMO, 40 ug/ml scrambled PMO (both synthesized by AVI-Biopharma),
repair vessel endothelium. We recently demonstrated that a transient blockade (2-4 days)
cells (Figure 2 & 6).
or medium alone then re-suspended in PBS alone for injection.
of endogenous transforming growth factor-beta type 1 (TGF-β1) in murine and human Ischemia/Reperfusion (I/R) assay: NOD-SCID mice (n = 6) were kept under Isoflurane vapor during induction of ischemia. The anterior chamber of the eye was 3) TGF-β PMO treatment of CD34+ cells from
hematopoietic stem cells (HSC) accelerates bone marrow engraftment, while dramatically cannulated using a 30-ga needle attached to an infusion line of saline and subjected to two hours of increased hydrostatic pressure in the anterior chamber. Retinal diabetic patients restores their defective
reducing the number of HSC needed for long-term reconstitution. TGF-β1 has been shown ischemia then ensued, confirmed by whitening of the iris and loss of the red reflux. After 2hrs. the needle was withdrawn and the intraocular pressure normalized,
migration and increases CXCR4 protein
resulting in reperfusion injury. The contra lateral eye served as a control. After 7 days retinal capillary damage was appreciable, and at that point human CD34+ cells
not only to downregulate stromal derived factor (SDF)-1, a key chemokine for EPC
expression (Figure 4).
from either diabetic or non-diabetic patients were injected intravitreally with 5000-20,000 cells/eye with either control PMO or TGF-β PMO. At 2-4 days the animals
chemotaxis, but also to down regulate the receptor for SDF-1 (CXCR4) on CD34+ HSC
4) TGF-β blockade dramatically increases the
were sacrificed, eyes enucleated, fixed in PFA and flat mounted. Following blocking in 10mM HEPES, 0.2 %BSA, 0.2 % Triton X, flat mounts were reacted overnight
and EPC, leading to detrimental effects on function of both cell types. Increasing CXCR4 with Rhodamine 1:1000 to visualize vasculature. The mounts were treated with primary human nuclear antigen antibodies in goat serum blocking buffer, followed by engraftment of human CD34+ stem cells into
on bone marrow HSC increases their homing to, and engraftment into the bone marrow, secondary staining with FITC-Goat-anti-mouse antibodies. Individual confocal images were analyzed using Image J and green fluorescence per 100 sq micron area per NOD-SCID mouse marrow (data not shown).
and into sites of ischemic and injured tissue where SDF-1 has been released. group. Data was analyzed using Graph pad with ANOVA followed by Tukey and plotted as Arbitrary Fluorescent Units” (AFU).
RESULTS SUMMARY
This study describes pre-clinical experimentation that may allow autologous human
CD34+ to be effectively and safely employed to treat diabetic retinopathy. No effective
treatment currently exists to reverse the vision loss resulting from retinal ischemia
(Fig A-C), Diabetic CD34+
associated with this condition. The chronic diabetic environment severely disrupts the
cells exposed to control
vascular repair capability of CD34+/EPC in the peripheral blood. However, patient
morpholino antisense PMO
CD34+ cells have the potential to generate large numbers of endothelial progenitor cells.
Here we show that blood derived diabetic CD34+ cells can repair damaged retinal
(Fig D-F), PMO targeted to
Figure 3: TGF-β, a pleiotropic autocrine and paracrine regulator of
TGF-β1, overnight, and vascular beds similar to non-diabetic CD34+ cells if TGF-β expression in the CD34+ cells
all stages of hematopoiesis is increased in HSC and serum of
patients with diabetes.
injected I/R mice
is transiently blocked.
Migration of human BM CD34+ Cells to SDF-1
(Fig G-I), Identical studies
were performed using human
Figure 2 80
REFERENCES
diabetic cells untreated
Migrating cells (RFU)
60
or treated with control PMO 40
20
1 Bartelmez SH, Iversen P, and Ruscetti FW. Blockade of endogenous TGFβ1 in hematopoietic stem cells
(Fig J-L), Identical studies 0
accelerates engraftment and enhances repopulating efficiency. Submitted to BLOOD 12/2008
Negative Medium PMO control PMO anti-
were performed using human -20 control alone TGF-β1 2 Ruscetti, F.W. & Bartelmez, S.H. Transforming growth factor beta, pleiotropic regulator of hematopoietic stem
diabetic cells treated with Figure 4: Migration of CD34+ cells toward SDF-1 increases after
cells: potential physiological and clinical relevance. Int J Hematol 74, 18-25. 2001.
control PMO TGF-β PMO treatment.
3 Harris JR, Brown GA, Jorgensen M, Kaushal S, Ellis EA, Grant MB, Scott EW. Bone marrow-derived cells
home to and regenerate retinal pigment epithelium after injury. Invest Ophthalmol Vis Sci. 2006
RPM I Scrambled TGF-be ta M AS
(Fig M-O), TGF-β1- PMO
May;47(5):2108-13.
200
180
4 Sengupta N, Caballero S, Mames RN, Butler JM, Scott EW, Grant MB. The role of adult bone marrow-derived
Fig 1:TGFß-1 blockade in HSC enhances their repair function 160
stem cells in choroidal neovascularization. Invest Ophthalmol Vis Sci. 2003 Nov;44(11):4908-13.
140
CXCR4 protein
120
5 Lanning DA, Nwomeh BC, Montante SJ, Yager DR, Diegelmann RF, Haynes JH. TGF-beta1 alters the healing
100
80
of cutaneous fetal excisional wounds. J Pediatr Surg. 1999 May;34(5):695-700.
Blood Vessels HSC in Murine Both 60
40
Figure 2 TGF-β PMO treatment of CD34+ cells from Diabetic and Non-diabetic patients 20
ASH Annual Meeting, December 6-9th 2008 Support: NIH EY07739, NIH EY12601, BetaStem Therapeutics Inc.
0
accelerates their vascular repair (I/R assay)
Commercial Relationship: S. Bartelmez & G. Gannon, BetaStem Therapeutics
Figure 5: CXCR4 Protein Expression Increases in
San Francisco, CA CD34+ after TGF-β PMO treatment