- LDOC1 mRNA expression is associated with markers of poor prognosis in chronic lymphocytic leukemia (CLL) and predicts overall survival better than other markers.
- A novel LDOC1 splice variant, LDOC1S, was identified that encodes only the leucine zipper domain. LDOC1S and LDOC1 are differentially expressed in normal and malignant B-cells.
- Knockdown of LDOC1 in HeLa cells using siRNA identified 122 differentially expressed genes involved in cancer pathways, suggesting LDOC1 may regulate genes important for proliferation and survival.
This document summarizes research on the immunobiology of chronic lymphocytic leukemia (CLL). Key points include:
1) CLL results from the transformation of B lymphocytes expressing antigen receptors (BCRs) of restricted structure that can be polyreactive and autoreactive.
2) These BCRs bind both natural antigens and novel autoantigens generated during apoptosis.
3) CLL subgroups differ in retention of polyreactivity, with retention associated with worse clinical outcomes.
This document summarizes research on the immunobiology of chronic lymphocytic leukemia (CLL). Key points include:
1. CLL results from the selection and transformation of B cells expressing B-cell receptors (BCRs) of restricted structure that can be polyreactive and autoreactive.
2. These BCRs bind to natural antigens as well as novel autoantigens generated during apoptosis.
3. CLL subgroups differ in retention of polyreactivity, with retention associated with worse clinical outcomes. Binding to autoantigens on apoptotic cells correlates with poor survival.
This document summarizes a study investigating the expression and prognostic significance of the LDOC1 gene in chronic lymphocytic leukemia (CLL) and normal B cells. The key findings are:
1) LDOC1 mRNA expression is significantly lower in CLL cases with mutated immunoglobulin variable region genes compared to unmutated cases.
2) LDOC1 is expressed in normal peripheral blood B cell subsets, suggesting it may play a role in normal B cell development.
3) High LDOC1 expression in CLL correlates with poor prognostic biomarkers and overall survival.
4) LDOC1 mRNA level is an excellent predictor of overall survival in untreated CLL patients.
Hematopoietic stem cell transplantation (HSCT) can cure thalassemia major by replacing diseased bone marrow. The process involves intensive preparation using chemotherapy and/or radiation to ablate the recipient's marrow. Healthy stem cells from a donor's bone marrow or umbilical cord blood are then infused. While HSCT from an HLA-identical sibling donor yields the best results, outcomes from unrelated donors are improving. Gene therapy clinical trials also aim to cure thalassemia by delivering healthy globin genes directly to the patient's stem cells.
1) Researchers identified differential methylation of the GRIN2D gene in colon cancer cells using reduced representation bisulfite sequencing, finding it demethylated in approximately 60% of cell lines. 2) They confirmed this epigenetic alteration led to upregulated GRIN2D expression and assessed its role in the cancer phenotype. 3) Knockdown of GRIN2D expression in demethylated RKO colon cancer cells using shRNA corresponded to reduced cell growth, suggesting GRIN2D demethylation contributes to the tumor cell phenotype. Future replication of these findings is needed to fully understand the significance.
This document describes a study that analyzed the frequency of cells with a PIG-A mutant phenotype (PNH phenotype) in samples from 19 patients with acute lymphoblastic leukemia (ALL) and immortalized B cell lines (BLCLs) from healthy donors. The BLCLs showed a median frequency of 11 x 10-6, while the ALL samples showed two distinct populations - about half of samples had a median frequency of 13 x 10-6, similar to controls, while the other half had a significantly higher median frequency of 566 x 10-6. This suggests that in ALL, there are two phenotypes with respect to hypermutability, which may correlate with the number of mutations required to cause the leukemia.
This document summarizes research on the major types of cell death: apoptosis, autophagy, and necrosis. It provides a brief history of the discovery and understanding of each type of cell death. It also describes the key morphological and molecular characteristics of each type of cell death, as well as physiological and pathophysiological functions. The document then discusses experimental approaches to study cell death mechanisms using tools from QIAGEN, including gene expression analysis, epigenetic analysis, and functional studies using reporter assays, siRNA, and shRNA.
New insights into leukemic niche in bone marrownanog
1. Leukemic stem cells home to and reside in the endosteal niche in the bone marrow, outcompeting normal hematopoietic stem cells. They rely on interactions with the microenvironment including secreted factors like SCF.
2. Neutralizing SCF disrupts the migration of normal hematopoietic progenitor cells to the leukemic niche and restores their mobilization by G-CSF, demonstrating SCF mediates the abnormal behavior of normal cells in the leukemic niche.
3. Targeting the SCF pathway may be a potential therapeutic strategy to disrupt the leukemic niche and improve outcomes for bone marrow transplantation and stem cell harvesting in leukemia patients.
This document summarizes research on the immunobiology of chronic lymphocytic leukemia (CLL). Key points include:
1) CLL results from the transformation of B lymphocytes expressing antigen receptors (BCRs) of restricted structure that can be polyreactive and autoreactive.
2) These BCRs bind both natural antigens and novel autoantigens generated during apoptosis.
3) CLL subgroups differ in retention of polyreactivity, with retention associated with worse clinical outcomes.
This document summarizes research on the immunobiology of chronic lymphocytic leukemia (CLL). Key points include:
1. CLL results from the selection and transformation of B cells expressing B-cell receptors (BCRs) of restricted structure that can be polyreactive and autoreactive.
2. These BCRs bind to natural antigens as well as novel autoantigens generated during apoptosis.
3. CLL subgroups differ in retention of polyreactivity, with retention associated with worse clinical outcomes. Binding to autoantigens on apoptotic cells correlates with poor survival.
This document summarizes a study investigating the expression and prognostic significance of the LDOC1 gene in chronic lymphocytic leukemia (CLL) and normal B cells. The key findings are:
1) LDOC1 mRNA expression is significantly lower in CLL cases with mutated immunoglobulin variable region genes compared to unmutated cases.
2) LDOC1 is expressed in normal peripheral blood B cell subsets, suggesting it may play a role in normal B cell development.
3) High LDOC1 expression in CLL correlates with poor prognostic biomarkers and overall survival.
4) LDOC1 mRNA level is an excellent predictor of overall survival in untreated CLL patients.
Hematopoietic stem cell transplantation (HSCT) can cure thalassemia major by replacing diseased bone marrow. The process involves intensive preparation using chemotherapy and/or radiation to ablate the recipient's marrow. Healthy stem cells from a donor's bone marrow or umbilical cord blood are then infused. While HSCT from an HLA-identical sibling donor yields the best results, outcomes from unrelated donors are improving. Gene therapy clinical trials also aim to cure thalassemia by delivering healthy globin genes directly to the patient's stem cells.
1) Researchers identified differential methylation of the GRIN2D gene in colon cancer cells using reduced representation bisulfite sequencing, finding it demethylated in approximately 60% of cell lines. 2) They confirmed this epigenetic alteration led to upregulated GRIN2D expression and assessed its role in the cancer phenotype. 3) Knockdown of GRIN2D expression in demethylated RKO colon cancer cells using shRNA corresponded to reduced cell growth, suggesting GRIN2D demethylation contributes to the tumor cell phenotype. Future replication of these findings is needed to fully understand the significance.
This document describes a study that analyzed the frequency of cells with a PIG-A mutant phenotype (PNH phenotype) in samples from 19 patients with acute lymphoblastic leukemia (ALL) and immortalized B cell lines (BLCLs) from healthy donors. The BLCLs showed a median frequency of 11 x 10-6, while the ALL samples showed two distinct populations - about half of samples had a median frequency of 13 x 10-6, similar to controls, while the other half had a significantly higher median frequency of 566 x 10-6. This suggests that in ALL, there are two phenotypes with respect to hypermutability, which may correlate with the number of mutations required to cause the leukemia.
This document summarizes research on the major types of cell death: apoptosis, autophagy, and necrosis. It provides a brief history of the discovery and understanding of each type of cell death. It also describes the key morphological and molecular characteristics of each type of cell death, as well as physiological and pathophysiological functions. The document then discusses experimental approaches to study cell death mechanisms using tools from QIAGEN, including gene expression analysis, epigenetic analysis, and functional studies using reporter assays, siRNA, and shRNA.
New insights into leukemic niche in bone marrownanog
1. Leukemic stem cells home to and reside in the endosteal niche in the bone marrow, outcompeting normal hematopoietic stem cells. They rely on interactions with the microenvironment including secreted factors like SCF.
2. Neutralizing SCF disrupts the migration of normal hematopoietic progenitor cells to the leukemic niche and restores their mobilization by G-CSF, demonstrating SCF mediates the abnormal behavior of normal cells in the leukemic niche.
3. Targeting the SCF pathway may be a potential therapeutic strategy to disrupt the leukemic niche and improve outcomes for bone marrow transplantation and stem cell harvesting in leukemia patients.
This document discusses the importance of choosing appropriate assays and models when studying hematopoietic stem cells (HSCs) and how results can be misleading if different assays and models are used. It summarizes several examples where different assays/models led to contradictory findings about leukemia stem cell phenotypes and the role of the Hedgehog signaling pathway in HSCs. The document advocates for using purified HSC populations and non-irradiated recipient mice when assessing HSC properties to avoid potential discrepancies between phenotype and function. It also discusses factors like cell variability, circadian rhythms and irradiation status of recipient mice that can influence variability in HSC assay results.
NSA Diagnostic Laboratory has been operating since 1958, founded by Prof. Nasseh Amin. NSA is considered as one of the most advanced labs in Egypt. Maintaining personalized services for its stakeholders, as well as the main role of the lab "Diagnosis"
NSA Diagnostic Laboratory operates through two different segments.
Firstly, a group of stand-alone labs located at prime locations all over Egypt, with the latest and up to date equipments.
Secondly, being the backbone of well reputed hospitals and some polyclinics where NSA is the lab that is responsible for all medical testing there, serving all our patients with class A quality.
Our main focus is delivering quality care and with Cost-value return. NSA plays a key role in improving the health of many Egyptians, by providing access to quality service for more than 200,000 patients annually.
This patient case involves a 56-year-old man with a history of JAK2 V617F+ essential thrombocythemia who developed severe anemia and was found to have myelodysplastic syndrome/myelofibrosis. He presented with transfusion-dependent anemia and was found to have concurrent warm autoantibody-mediated hemolytic anemia and delayed hemolytic transfusion reaction due to alloimmunization to the Kell blood group antigen. He required intensive care for management of his conditions.
This study found that oncogenic Ras sensitizes normal human cells to TRAIL-induced apoptosis by enhancing caspase 8 recruitment and activation at the DISC. Specifically:
1) Normal and immortalized human cells were resistant to TRAIL, while Ras-transformed cells were susceptible, undergoing apoptosis.
2) Ras transformation potentiated TRAIL-induced cleavage of caspase 8 and its substrates Bid and plectin, indicating enhanced caspase 8 activation.
3) Ras enhanced the recruitment of caspase 8 to the TRAIL death-inducing signaling complex (DISC), allowing more efficient caspase 8 cleavage and activation of the apoptotic pathway.
AACR 2013 Abstract 2047 Poster - SLITRK6 in Bladder Cancer - YSDeanna Russell
AGS-15/SLITRK6 was discovered as a differentially expressed gene in bladder cancer. It encodes a cell surface protein with neuronal functions and restricted normal tissue expression. High and frequent expression of SLITRK6 was found in bladder cancers and other cancer types. Patient-derived xenografts and cancer cell lines with SLITRK6 expression were identified as models for antibody therapeutic development targeting SLITRK6, particularly for bladder cancer. Comprehensive analysis revealed SLITRK6's selective expression in tumors makes it a suitable candidate for antibody-drug conjugate cancer treatment.
Transplantation in sensitized patients(seminar)Vishal Golay
This document discusses HLA typing, crossmatching, and transplantation in sensitized patients. It begins with a brief history of organ transplantation. It then covers topics such as the structure and function of MHC molecules, methods of HLA typing including serological and DNA-based techniques, interpreting HLA typing reports, detecting sensitization through antibody detection tests, defining and identifying sensitized patients, and challenges in transplanting sensitized patients. Advances in diagnostics and therapeutics have helped increase transplantation options for sensitized patients.
Cord blood is a naturally discarded tissue that contains a high proportion of circulating hematopoietic stem cells. It allows HLA mismatch transplantation, providing cure to hematological malignancies with an attenuated risk of severe graft-versus-host disease through a graft-versus-leukemia effect. Cord blood transplantation is unique in its immunogenetics profile, preserving the immune cells carried in the cord blood graft, including T-regulatory cells and natural killer cells. Early reconstitution of lymphocytes from cord blood grafts predicts better outcomes. Advances in cord blood transplantation aim to take advantage of its immunological properties through donor selection accounting for HLA matching, maternal antigens, and cell dose to maximize graft-versus-leuke
1. The document analyzes the expression and activity of drug efflux pumps ABCB1 and ABCC1 in acute myeloid leukemia (AML) cell lines, xenografts, and patient samples.
2. It finds that ABCB1 expression and activity confers resistance to chemotherapies that are substrates of ABCB1 in AML cell lines and xenograft models.
3. ABCC1 expression is also detected in AML models and may contribute to drug resistance, though additional investigation is needed to understand its role.
ABSTRACT- Coronary artery disease (CAD) is suspected as a leading cause of mortality in developed countries. Due
to cholesterol and fat deposit plaque is forming into the inner walls of the arteries of the heart, which leads to narrowing
of blood vessels of heart and reduce the blood flow rate into heart. Proprotein convertase subtilisin-like kexin type 9
(PCSK9) is one of the candidate gene that regulate lipoprotein retention pathway of CAD development. It is a newly
discovered serine protease that plays a key role in LDL-C homeostasis by mediating LDL receptor (LDLR). The LDL
receptor is breakdown through a post transcriptional mechanism and induces the production of very low-density
lipoprotein in the fasting state. The aim of this study was to investigate the frequency of single nucleotide
polymorphism (SNP) of PCSK9 gene of 155 CAD patients and 102 ages matched healthy controls. Serum lipids
including total cholesterol (TC), triglycerides (TG), HDL, LDL, and VLDL were analyzed. PCR-RFLP analysis was
carried out to genotype regions carrying Eam 1104I restriction site in the PCSK9. Gene considering significant
difference in serum TC, TG, HDL-C, LDL-C and VLDL-C levels (P<0.001, <0.0001) of patients and control samples.
In CAD patients, G allele frequency is less than A allele frequency. G allele is responsible for decreasing the
LDL: HDL ratio which shows evidence in having its protecting effect on the occurrence of CAD in West Bengal Population.
Key-words- CAD, PCSK9, SNP, Eam1104I, Polymorphism, West Bengal population
Cdc6 Knockdown Renders p16INK4a Re-Activation, Leading to Senescence Human Br...gan-navi
Luo Feng, Jerry R. Barnhart, Lingtao Wu, Greg Shackleford,
Sheng-he Huang and Ambrose Jong
Department of Hematology and Oncology
Childrens Hospital Los Angeles
University of Southern California, Los Angeles
USA
The document describes a study that investigated oxidative stress and cell death in lymphoblastoid cell lines (LCLs) from children with autism as compared to controls. The study found that LCLs from children with autism exhibited higher baseline levels of reactive oxygen species and were more susceptible to oxidative stress induced by DMNQ treatment. Autistic LCLs showed greater changes in cell viability and higher rates of cell death compared to controls after DMNQ treatment. Control LCLs exhibited higher levels of apoptosis, while autistic LCLs showed more necrosis. The results suggest children with autism have an impaired ability to combat oxidative stress that may be related to lower glutathione levels.
Cellular Immune Therapy with Allogeneic Stem Cell Transplantationspa718
This document discusses cellular immune therapy approaches using hematopoietic stem cell transplantation. It notes that high-dose chemotherapy alone does not fully eradicate malignancies and relapse remains a major treatment failure. The graft-versus-malignancy effect provided by allogeneic transplantation is responsible for residual disease eradication but is associated with graft-versus-host disease. The document reviews approaches to enhance the graft-versus-malignancy effect through donor lymphocyte infusions, antigen-specific cytotoxic T-cells, and chimeric antigen receptor T-cells while preventing graft-versus-host disease using regulatory T-cells or genetically modified T-cells with suicide switches. Ongoing clinical trials
This study investigates the interaction between splicing and transcriptional elongation in vivo using a zebrafish mutant for the spliceosomal component sf3b1. The researchers conducted Pol II ChIP-seq on wild type and mutant embryos to detect transcriptional stalling, indicated by additional Pol II binding sites along gene bodies in mutants. They also measured RNA synthesis in endothelial cells, finding a two-fold decrease in mutants, indicating reduced transcriptional output. Future work will further examine nascent RNA and identify endothelial subpopulations with high transcription relevant to hematopoietic stem cell development. The research aims to characterize how spliceosome disruption affects transcription using zebrafish as an in vivo model.
This document discusses phenotypic heterogeneity in multiple myeloma and identifies distinct subclones within patient samples that have different genomic profiles, clonogenic potential, and drug sensitivity. FACS-sorted subclones from patient samples are often associated with different cytogenetic profiles. After drug exposure, minimal residual disease is a reservoir of chemoresistant cells that can lead to relapse. Achieving a complete response and eliminating minimal residual disease is associated with longer survival times.
Clinical diagnosis of chronic myeloid leukemia by real time polymerase chain ...Teboho Mooko
Oncology study i did in my third year (2014). the study was basically about monitoring Chronic Myeloid leukemia (CML) using Real-Time PCR techniques to check how patients from Universitas Hospital responded to the treatment of Gleevec drug.
Cytogenetic Analysis in Hematological Malignanciesspa718
The document discusses the importance of cytogenetic analysis in hematological malignancies. Some key points:
- Many hematological malignancies have clonal chromosomal abnormalities that can aid in diagnosis, classification, and risk stratification.
- Certain recurrent abnormalities are specific to certain tumor subtypes and can predict treatment response and clinical outcome.
- Genes at breakpoints of recurrent abnormalities play a role in tumorigenesis and can be treatment targets.
- Cytogenetic analysis is useful for diagnosis, risk stratification, treatment selection, and monitoring treatment response in hematological cancers like CML, AML, ALL, lymphoma, MDS, MM, and CLL.
Pells et al [2015] PLoS ONE 10[7] e0131102Steve Pells
This research article identifies novel human embryonic stem cell regulators based on their conserved and distinct CpG island methylation patterns. The researchers analyzed CpG island methylation in four human embryonic stem cell lines using a CpG island array and identified 1,111 CpGs that were methylated in all stem cell lines. They compared the methylation profiles to somatic tissues and mRNA expression data to identify stem cell-specific methylation patterns associated with gene expression. Genes related to transcriptional repression and activation were overrepresented among genes associated with methylated or unmethylated CpGs specifically in stem cells. Knockdown experiments confirmed that some candidate regulators induced stem cell differentiation, while overexpression modulated induced pluripotent stem cell formation
Cdc6 knockdown inhibits human neuroblastoma cell proliferationgan-navi
Luo Feng Æ Jerry R. Barnhart Æ Robert C. Seeger Æ Lingtao Wu Æ
Nino Keshelava Æ Sheng-He Huang Æ Ambrose Jong
Received: 19 October 2007 / Accepted: 10 January 2008
Springer Science+Business Media, LLC. 2008
This document summarizes key information about chronic lymphocytic leukemia (CLL) including its invisible, inconclusive, and incurable nature at diagnosis. It notes that CLL is often asymptomatic at diagnosis but can have an aggressive clinical course over time, reflecting underlying biological heterogeneity. While treatment has progressed, CLL remains incurable. The document discusses CLL diagnosis and treatment timing and changes in the host and tumor over time. It provides background on CLL occurring in mature B cells and the importance of signaling pathways. The document summarizes prognostic factors in CLL including genetics, mutations, and biomarkers from the tumor and microenvironment. It notes the importance of biological risk stratification and targeting signaling pathways in CLL treatment.
Verifying the role of AID in Chronic Lymphocytic LeukemiaCharlotte Broadbent
This study aimed to verify the role of the enzyme activation-induced deaminase (AID) in chronic lymphocytic leukemia (CLL) using statistical bootstrapping methods. DNA sequences from CLL patients were analyzed before and after AID stimulation. Three tests found statistically significant evidence that AID is active in CLL: 1) more mutations in the variable region than constant region, 2) more mutations at AID hotspots than expected, and 3) fewer mutations at AID coldspots than expected. The results confirm AID's role in somatic hypermutation in CLL, furthering understanding of this disease.
Steven M. Kornblau is a professor of medicine at UT MD Anderson Cancer Center who specializes in hematology and oncology. He completed his medical training at MD Anderson from 1988-1991 and has been a faculty member there since 1991. Dr. Kornblau's research focuses on protein expression patterns in blood cancers. He also runs one of the largest tissue repositories for leukemia and myeloma in the world. The document discusses myelofibrosis, its diagnosis, prognosis, driver mutations, complications, and traditional and newer treatment options like ruxolitinib.
Ohio State's ASH Review 2017 - Myeloproliferative DisordersOSUCCC - James
Katherine Walsh, MD
Assistant Professor of Clinical Internal Medicine
The Ohio State University Comprehensive Cancer Center -
Arthur G. James Cancer Hospital and Richard J. Solove Research Institute
This document discusses the importance of choosing appropriate assays and models when studying hematopoietic stem cells (HSCs) and how results can be misleading if different assays and models are used. It summarizes several examples where different assays/models led to contradictory findings about leukemia stem cell phenotypes and the role of the Hedgehog signaling pathway in HSCs. The document advocates for using purified HSC populations and non-irradiated recipient mice when assessing HSC properties to avoid potential discrepancies between phenotype and function. It also discusses factors like cell variability, circadian rhythms and irradiation status of recipient mice that can influence variability in HSC assay results.
NSA Diagnostic Laboratory has been operating since 1958, founded by Prof. Nasseh Amin. NSA is considered as one of the most advanced labs in Egypt. Maintaining personalized services for its stakeholders, as well as the main role of the lab "Diagnosis"
NSA Diagnostic Laboratory operates through two different segments.
Firstly, a group of stand-alone labs located at prime locations all over Egypt, with the latest and up to date equipments.
Secondly, being the backbone of well reputed hospitals and some polyclinics where NSA is the lab that is responsible for all medical testing there, serving all our patients with class A quality.
Our main focus is delivering quality care and with Cost-value return. NSA plays a key role in improving the health of many Egyptians, by providing access to quality service for more than 200,000 patients annually.
This patient case involves a 56-year-old man with a history of JAK2 V617F+ essential thrombocythemia who developed severe anemia and was found to have myelodysplastic syndrome/myelofibrosis. He presented with transfusion-dependent anemia and was found to have concurrent warm autoantibody-mediated hemolytic anemia and delayed hemolytic transfusion reaction due to alloimmunization to the Kell blood group antigen. He required intensive care for management of his conditions.
This study found that oncogenic Ras sensitizes normal human cells to TRAIL-induced apoptosis by enhancing caspase 8 recruitment and activation at the DISC. Specifically:
1) Normal and immortalized human cells were resistant to TRAIL, while Ras-transformed cells were susceptible, undergoing apoptosis.
2) Ras transformation potentiated TRAIL-induced cleavage of caspase 8 and its substrates Bid and plectin, indicating enhanced caspase 8 activation.
3) Ras enhanced the recruitment of caspase 8 to the TRAIL death-inducing signaling complex (DISC), allowing more efficient caspase 8 cleavage and activation of the apoptotic pathway.
AACR 2013 Abstract 2047 Poster - SLITRK6 in Bladder Cancer - YSDeanna Russell
AGS-15/SLITRK6 was discovered as a differentially expressed gene in bladder cancer. It encodes a cell surface protein with neuronal functions and restricted normal tissue expression. High and frequent expression of SLITRK6 was found in bladder cancers and other cancer types. Patient-derived xenografts and cancer cell lines with SLITRK6 expression were identified as models for antibody therapeutic development targeting SLITRK6, particularly for bladder cancer. Comprehensive analysis revealed SLITRK6's selective expression in tumors makes it a suitable candidate for antibody-drug conjugate cancer treatment.
Transplantation in sensitized patients(seminar)Vishal Golay
This document discusses HLA typing, crossmatching, and transplantation in sensitized patients. It begins with a brief history of organ transplantation. It then covers topics such as the structure and function of MHC molecules, methods of HLA typing including serological and DNA-based techniques, interpreting HLA typing reports, detecting sensitization through antibody detection tests, defining and identifying sensitized patients, and challenges in transplanting sensitized patients. Advances in diagnostics and therapeutics have helped increase transplantation options for sensitized patients.
Cord blood is a naturally discarded tissue that contains a high proportion of circulating hematopoietic stem cells. It allows HLA mismatch transplantation, providing cure to hematological malignancies with an attenuated risk of severe graft-versus-host disease through a graft-versus-leukemia effect. Cord blood transplantation is unique in its immunogenetics profile, preserving the immune cells carried in the cord blood graft, including T-regulatory cells and natural killer cells. Early reconstitution of lymphocytes from cord blood grafts predicts better outcomes. Advances in cord blood transplantation aim to take advantage of its immunological properties through donor selection accounting for HLA matching, maternal antigens, and cell dose to maximize graft-versus-leuke
1. The document analyzes the expression and activity of drug efflux pumps ABCB1 and ABCC1 in acute myeloid leukemia (AML) cell lines, xenografts, and patient samples.
2. It finds that ABCB1 expression and activity confers resistance to chemotherapies that are substrates of ABCB1 in AML cell lines and xenograft models.
3. ABCC1 expression is also detected in AML models and may contribute to drug resistance, though additional investigation is needed to understand its role.
ABSTRACT- Coronary artery disease (CAD) is suspected as a leading cause of mortality in developed countries. Due
to cholesterol and fat deposit plaque is forming into the inner walls of the arteries of the heart, which leads to narrowing
of blood vessels of heart and reduce the blood flow rate into heart. Proprotein convertase subtilisin-like kexin type 9
(PCSK9) is one of the candidate gene that regulate lipoprotein retention pathway of CAD development. It is a newly
discovered serine protease that plays a key role in LDL-C homeostasis by mediating LDL receptor (LDLR). The LDL
receptor is breakdown through a post transcriptional mechanism and induces the production of very low-density
lipoprotein in the fasting state. The aim of this study was to investigate the frequency of single nucleotide
polymorphism (SNP) of PCSK9 gene of 155 CAD patients and 102 ages matched healthy controls. Serum lipids
including total cholesterol (TC), triglycerides (TG), HDL, LDL, and VLDL were analyzed. PCR-RFLP analysis was
carried out to genotype regions carrying Eam 1104I restriction site in the PCSK9. Gene considering significant
difference in serum TC, TG, HDL-C, LDL-C and VLDL-C levels (P<0.001, <0.0001) of patients and control samples.
In CAD patients, G allele frequency is less than A allele frequency. G allele is responsible for decreasing the
LDL: HDL ratio which shows evidence in having its protecting effect on the occurrence of CAD in West Bengal Population.
Key-words- CAD, PCSK9, SNP, Eam1104I, Polymorphism, West Bengal population
Cdc6 Knockdown Renders p16INK4a Re-Activation, Leading to Senescence Human Br...gan-navi
Luo Feng, Jerry R. Barnhart, Lingtao Wu, Greg Shackleford,
Sheng-he Huang and Ambrose Jong
Department of Hematology and Oncology
Childrens Hospital Los Angeles
University of Southern California, Los Angeles
USA
The document describes a study that investigated oxidative stress and cell death in lymphoblastoid cell lines (LCLs) from children with autism as compared to controls. The study found that LCLs from children with autism exhibited higher baseline levels of reactive oxygen species and were more susceptible to oxidative stress induced by DMNQ treatment. Autistic LCLs showed greater changes in cell viability and higher rates of cell death compared to controls after DMNQ treatment. Control LCLs exhibited higher levels of apoptosis, while autistic LCLs showed more necrosis. The results suggest children with autism have an impaired ability to combat oxidative stress that may be related to lower glutathione levels.
Cellular Immune Therapy with Allogeneic Stem Cell Transplantationspa718
This document discusses cellular immune therapy approaches using hematopoietic stem cell transplantation. It notes that high-dose chemotherapy alone does not fully eradicate malignancies and relapse remains a major treatment failure. The graft-versus-malignancy effect provided by allogeneic transplantation is responsible for residual disease eradication but is associated with graft-versus-host disease. The document reviews approaches to enhance the graft-versus-malignancy effect through donor lymphocyte infusions, antigen-specific cytotoxic T-cells, and chimeric antigen receptor T-cells while preventing graft-versus-host disease using regulatory T-cells or genetically modified T-cells with suicide switches. Ongoing clinical trials
This study investigates the interaction between splicing and transcriptional elongation in vivo using a zebrafish mutant for the spliceosomal component sf3b1. The researchers conducted Pol II ChIP-seq on wild type and mutant embryos to detect transcriptional stalling, indicated by additional Pol II binding sites along gene bodies in mutants. They also measured RNA synthesis in endothelial cells, finding a two-fold decrease in mutants, indicating reduced transcriptional output. Future work will further examine nascent RNA and identify endothelial subpopulations with high transcription relevant to hematopoietic stem cell development. The research aims to characterize how spliceosome disruption affects transcription using zebrafish as an in vivo model.
This document discusses phenotypic heterogeneity in multiple myeloma and identifies distinct subclones within patient samples that have different genomic profiles, clonogenic potential, and drug sensitivity. FACS-sorted subclones from patient samples are often associated with different cytogenetic profiles. After drug exposure, minimal residual disease is a reservoir of chemoresistant cells that can lead to relapse. Achieving a complete response and eliminating minimal residual disease is associated with longer survival times.
Clinical diagnosis of chronic myeloid leukemia by real time polymerase chain ...Teboho Mooko
Oncology study i did in my third year (2014). the study was basically about monitoring Chronic Myeloid leukemia (CML) using Real-Time PCR techniques to check how patients from Universitas Hospital responded to the treatment of Gleevec drug.
Cytogenetic Analysis in Hematological Malignanciesspa718
The document discusses the importance of cytogenetic analysis in hematological malignancies. Some key points:
- Many hematological malignancies have clonal chromosomal abnormalities that can aid in diagnosis, classification, and risk stratification.
- Certain recurrent abnormalities are specific to certain tumor subtypes and can predict treatment response and clinical outcome.
- Genes at breakpoints of recurrent abnormalities play a role in tumorigenesis and can be treatment targets.
- Cytogenetic analysis is useful for diagnosis, risk stratification, treatment selection, and monitoring treatment response in hematological cancers like CML, AML, ALL, lymphoma, MDS, MM, and CLL.
Pells et al [2015] PLoS ONE 10[7] e0131102Steve Pells
This research article identifies novel human embryonic stem cell regulators based on their conserved and distinct CpG island methylation patterns. The researchers analyzed CpG island methylation in four human embryonic stem cell lines using a CpG island array and identified 1,111 CpGs that were methylated in all stem cell lines. They compared the methylation profiles to somatic tissues and mRNA expression data to identify stem cell-specific methylation patterns associated with gene expression. Genes related to transcriptional repression and activation were overrepresented among genes associated with methylated or unmethylated CpGs specifically in stem cells. Knockdown experiments confirmed that some candidate regulators induced stem cell differentiation, while overexpression modulated induced pluripotent stem cell formation
Cdc6 knockdown inhibits human neuroblastoma cell proliferationgan-navi
Luo Feng Æ Jerry R. Barnhart Æ Robert C. Seeger Æ Lingtao Wu Æ
Nino Keshelava Æ Sheng-He Huang Æ Ambrose Jong
Received: 19 October 2007 / Accepted: 10 January 2008
Springer Science+Business Media, LLC. 2008
This document summarizes key information about chronic lymphocytic leukemia (CLL) including its invisible, inconclusive, and incurable nature at diagnosis. It notes that CLL is often asymptomatic at diagnosis but can have an aggressive clinical course over time, reflecting underlying biological heterogeneity. While treatment has progressed, CLL remains incurable. The document discusses CLL diagnosis and treatment timing and changes in the host and tumor over time. It provides background on CLL occurring in mature B cells and the importance of signaling pathways. The document summarizes prognostic factors in CLL including genetics, mutations, and biomarkers from the tumor and microenvironment. It notes the importance of biological risk stratification and targeting signaling pathways in CLL treatment.
Verifying the role of AID in Chronic Lymphocytic LeukemiaCharlotte Broadbent
This study aimed to verify the role of the enzyme activation-induced deaminase (AID) in chronic lymphocytic leukemia (CLL) using statistical bootstrapping methods. DNA sequences from CLL patients were analyzed before and after AID stimulation. Three tests found statistically significant evidence that AID is active in CLL: 1) more mutations in the variable region than constant region, 2) more mutations at AID hotspots than expected, and 3) fewer mutations at AID coldspots than expected. The results confirm AID's role in somatic hypermutation in CLL, furthering understanding of this disease.
Steven M. Kornblau is a professor of medicine at UT MD Anderson Cancer Center who specializes in hematology and oncology. He completed his medical training at MD Anderson from 1988-1991 and has been a faculty member there since 1991. Dr. Kornblau's research focuses on protein expression patterns in blood cancers. He also runs one of the largest tissue repositories for leukemia and myeloma in the world. The document discusses myelofibrosis, its diagnosis, prognosis, driver mutations, complications, and traditional and newer treatment options like ruxolitinib.
Ohio State's ASH Review 2017 - Myeloproliferative DisordersOSUCCC - James
Katherine Walsh, MD
Assistant Professor of Clinical Internal Medicine
The Ohio State University Comprehensive Cancer Center -
Arthur G. James Cancer Hospital and Richard J. Solove Research Institute
Translational Genomics and Prostate Cancer: Meet the NGS Experts Series Part 2QIAGEN
Advanced prostate cancer is highly heterogeneous but this inter-patient heterogeneity has until recently not been understood. We have through an international research effort dissected the molecular landscape of advanced castration resistant prostate, elucidating key molecular targets in this group of diseases. We have also shown that PARP inhibitors have antitumor activity against a significant proportion of these cancers, mainly in men whose cancers harbor DNA repair defects.
Chronic lymphocytic leukemia (CLL) is characterized by the proliferation and accumulation of small, mature lymphocytes in the blood, bone marrow, and lymphoid tissues. CLL is diagnosed based on an absolute lymphocyte count over 5000 with immunophenotyping showing a clonal CD5+/CD19+/CD23+ B-cell population. Prognosis is based on factors like clinical stage, bone marrow histology, lymphocyte doubling time, genetic abnormalities, CD38 and ZAP-70 expression levels, and IgVH mutation status. Treatment options range from watchful waiting to chemotherapy, chemoimmunotherapy, monoclonal antibodies, and stem cell transplantation depending on prognostic factors and symptom severity.
This document discusses acute leukemia, including its definition, predisposing factors, mechanisms, clinical features, laboratory diagnosis, and WHO classification. Acute leukemia originates from hematopoietic stem cells and is characterized by proliferation of immature blast cells in the bone marrow. It comprises about 50% of leukemic cases and can be either acute lymphoblastic leukemia (ALL) or acute myeloid leukemia (AML). Diagnosis involves morphological examination of blood and bone marrow smears, cytochemistry, immunophenotyping, cytogenetics, and molecular studies to classify it as ALL or AML and identify specific genetic abnormalities. Prognosis and treatment depend on identified risk factors and mutations.
The document summarizes research on targeting cancer stem cells in acute myeloid leukemia (AML). It discusses how AML stem cells, or leukemia stem cells (LSCs), drive leukemia but are overlooked by standard treatment. The research aims to understand LSC function at a molecular level to enable LSC-directed therapies for AML cure. Experiments show the transcription factor PU.1 regulates LSCs, and the drug eltrombopag may help treat thrombocytopenia in AML without activating LSCs.
The document summarizes research on targeting cancer stem cells in acute myeloid leukemia (AML). It discusses how AML stem cells, or leukemia stem cells (LSCs), differ from normal stem cells and bulk leukemia cells in their ability to self-renew and initiate leukemia. The research aims to identify molecular mechanisms driving LSC function and transforming events leading to LSC formation. Experiments show that reducing the transcription factor PU.1 induces AML by downregulating JunB expression. Additional work profiles gene expression differences between normal and AML stem/progenitor cells. The document also evaluates using the thrombopoietin receptor agonist eltrombopag to treat thrombocytopenia in AML/MDS patients,
The document summarizes research on targeting cancer stem cells in acute myeloid leukemia (AML). It discusses how AML stem cells, or leukemia stem cells (LSCs), differ from normal stem cells and bulk leukemia cells in their ability to self-renew and initiate leukemia. The research aims to identify molecular mechanisms driving LSC function and transforming events leading to LSC formation. Experiments show that reducing the transcription factor PU.1 induces AML by downregulating JunB expression. Additional work profiles gene expression differences between normal and AML stem/progenitor cells. The document also evaluates using the thrombopoietin receptor agonist eltrombopag to treat thrombocytopenia in AML/MDS patients,
The document summarizes research on targeting cancer stem cells in acute myeloid leukemia (AML). It discusses how AML stem cells (leukemia stem cells or LSCs) differ from normal stem cells in their ability to self-renew and initiate leukemia. The research aims to understand the molecular mechanisms that drive LSC function and identify transforming events involved in LSC formation and maintenance. Preclinical studies show that the transcription factor PU.1 regulates genes like JunB that are important for LSC function. The document also discusses using a small molecule drug called Eltrombopag to stimulate megakaryopoiesis in MDS/AML patients without activating LSCs, as a potential treatment for thrombocytopenia.
This document summarizes clinical trial results of the oral ALK inhibitor crizotinib (PF-02341066) in patients with ALK-positive non-small cell lung cancer. The trial showed that crizotinib had significant clinical activity in ALK-positive NSCLC patients, with an objective response rate of 57% and disease control rate of 87%. The median progression-free survival was 10 months for these patients. In contrast, patients with ALK-negative NSCLC did not respond to crizotinib. These results provide evidence that crizotinib is an effective targeted therapy for patients with ALK-positive NSCLC.
Mahra Nourbakhsh's presentation, Hepatitis C Virus #1Mahra Nourbakhsh
The document summarizes information about hepatitis C virus (HCV) infection globally and in Canada. It provides statistics showing that an estimated 169.7 million people worldwide have HCV infection, with the highest prevalence rates in Africa, Southeast Asia, and the Eastern Mediterranean. In Canada, an estimated 242,500 people have HCV infection, with 7,900 new cases estimated in 2007. The document then outlines various aspects of HCV including transmission routes, natural history, diagnostic tools, viral life cycle, immunopathogenesis, progression of liver disease, and potential mechanisms of hepatic steatosis associated with HCV.
This document provides an overview and agenda for a discussion on diffuse large B-cell lymphoma (DLBCL) and its current treatment landscape. The agenda includes: initial management of limited-stage and advanced-stage DLBCL, predicting risk of central nervous system recurrence, and ongoing trials. Current standard first-line treatment for advanced DLBCL is R-CHOP chemotherapy. Recent trials have explored reducing chemotherapy cycles or adding targeted therapies such as polatuzumab vedotin to improve outcomes. Ongoing research focuses on risk stratification and personalized treatment approaches.
This document summarizes a study on kinase-activating lesions in Philadelphia chromosome-like (Ph-like) acute lymphoblastic leukemia (ALL). The study found that 91% of Ph-like ALL cases had genetic alterations activating kinase signaling, with subgroups involving kinases like ABL1, CSF1R, PDGFRB, EPOR, JAK2, CRLF2 and others. Patients with Ph-like ALL had poorer outcomes than those without, and the presence of alterations like IKZF1 deletions conferred even worse prognosis. In vitro experiments showed that fusions involving ABL1, ABL2 and CSF1R were sensitive to tyrosine kinase inhibitors like imatinib and dasatinib.
Newer biomarkers,techniques & their inclusion in 2016 WHO classification for leukaemia/lymphomas increases the responsibility of the pathologists, requiring to develop an integrated multidisciplinary approach for reporting.
NSCLC: diagnóstico molecular, pronóstico y seguimiento; CTCMauricio Lema
Lo nuevo en diagnóstico molecular, pronóstico y seguimiento en NSCLC, y el impacto pronóstico de las Células Tumorales Circulantes. Para evento de cirugía de tórax, Hotel Intercontinental, Medellín, 22.05.2018 (se complementa con las la presentación de lo nuevo en terapia sistémica en NSCLC).
This document summarizes studies evaluating immune checkpoint inhibitors for the treatment of Hodgkin lymphoma. It discusses pivotal trials that led to FDA approval of nivolumab and pembrolizumab for relapsed/refractory HL after stem cell transplant and brentuximab vedotin. It also reviews mechanisms of action of PD-1 blockade in HL and efforts to combine checkpoint inhibitors with other agents or incorporate them into frontline treatment for high-risk patients.
This study analyzed mutations in the von Hippel-Lindau (VHL) gene in 67 cases of renal cell carcinoma (RCC) to determine if VHL mutations could help distinguish between RCC subtypes. Single strand conformational polymorphism analysis and sequencing identified VHL mutations in RCC samples. Exon 3 mutations were found primarily in conventional (clear cell) RCC and correlated with more aggressive tumor phenotypes. Mutations were absent in papillary RCC and oncocytoma samples. VHL mutations were found to help distinguish conventional RCC from chromophobe RCC showing clear cells, serving as an adjunct to histological diagnosis of RCC subtypes.
TREATMENT OF NON-HIDGKIN'S LYMPHOMA IN ELDERLY PATIENTSspa718
This document summarizes treatment approaches for non-Hodgkin's lymphoma in elderly patients. It discusses palliative options for refractory/relapsed diffuse large B-cell lymphoma such as gemcitabine-based chemotherapy, low-dose oral chemotherapy, and hyperfractionated cyclophosphamide. It also reviews novel anti-CD20 monoclonal antibodies showing efficacy against relapsed/refractory indolent lymphoma, and brentuximab vedotin's mechanism of action and responses seen in relapsed/refractory systemic anaplastic large cell lymphoma. Finally, it provides a high-level overview of the MD Anderson Cancer Center's Department of Lymphoma/Myeloma and its disease-specific
TREATMENT OF NON-HIDGKIN'S LYMPHOMA IN ELDERLY PATIENTS
PhD-Defense-May 13 2011
1. LDOC1, a Novel Biomarker of Prognosis
in Chronic Lymphocytic Leukemia (CLL)
Hatice Duzkale
UT Graduate School of Biomedical Sciences at Houston
UT MD Anderson Cancer Center
Department of Hematopathology
Ph.D. Defense
May 16, 2011
2. Chronic Lymphocytic Leukemia
(CLL)
-most common leukemia
-disease of advanced age
-unknown etiology
-no curative treatment
-unpredictable prognosis
-1/3 indolent disease with normal survival
-1/3 indolent initially, progresses
-1/3 aggressive disease
3. Chronic Lymphocytic Leukemia
Prognostic Subtypes
Aggressive CLLIndolent CLL
Good Prognosis
Low Rai stage
Low β2-microglobulin
Low ZAP70
Del 13q
Mutated IGHV
Poor Prognosis
High Rai stage
High β2-microglobulin
High ZAP70
Del 11q, del 17p, tris 12
Unmutated IGHV
IGHV: Immunoglobulin Heavy chain Variable region
4. Immunoglobulin Somatic Mutation is
a Physiologic Process
modified from Kuppers et al., Nature Reviews Cancer, 2005, 5:251-262
DNA modifications
5. IGHV Mutation Status Predicts Survival
Median survival: 8yr vs 25yr
8yr 15yr 25yr
Unmutated IGHV
Mutated IGHV
Hamblin TJ, et al Blood. 1999 94(6):1848-54
6. Disease Subtype Classification
by IGHV Somatic Mutation Status
Novel Biomarkers Novel Molecular Targets
Global Gene Expression Profiling
8. LDOC1 Distinguishes CLL Prognostic Subtypes,
Based on IGHV Mutation Status
Abruzzo LV et al., J Mol Diagn. 2007, 9 (4):546-555
Screening of CLL literature for microarray studies
for genes distinguishing
unmutated from mutated CLL
Evaluation of 88 genes in
49 untreated CLL patients
(MF-QRT-PCR)
36 genes significant
LDOC1 most significant
unmutated CLL, mutated CLL
9. Can LDOC1 Predict Survival?
Evaluation of LDOC1 in 131 untreated CLL patients
(MF-QRT-PCR)
Correlation with
other prognostic parameters
Survival analysis
14. LDOC1 mRNA Expression Predicts Overall Survival
Better than IGHV Somatic Mutation Status and ZAP70
ZAP70IGHV
LDOC1
Multivariate
Analysis
Duzkale H. et al., Blood 2011, 117(15):4076-84
15. -LDOC1 mRNA expression is associated with markers
of poor prognosis in CLL
-unmutated IGHV status
-high ZAP70 protein expression
-cytogenetic abnormalities
-LDOC1 mRNA predicts overall survival
-LDOC1 mRNA predicts overall survival better than
IGHV mutation status and ZAP70 protein
LDOC1 mRNA Expression as a Biomarker
-Summary
16. Ellenberger et al., 1992
-putative Leucine zipper transcription factor
-ubiquitously expressed in normal tissues
-differentially expressed in cancer cell lines
- pancreatic and gastric cancer
- breast cancer
-anti-proliferative?
-pro-apoptotic?
-reduces NF-kB activity?
LDOC1
(Leucine Zipper Downregulated in Cancer)
20. LDOC1S is a Leucine zipper region alone
CDS!5’UTR! 3’UTR! CDS!5’UTR! 3’UTR!
Proline-rich
region!
Acidic
region!
Leucine
zipper!
Leucine
zipper!
Ellenberger et al., 1992
Splice Variant transcript
(LDOC1S)
Wild Type LDOC1 transcript
(LDOC1)
LDOC1S NCBI accession numbers
mRNA: HQ343285
Protein: ADO32619
21. How much of the total LDOC1 mRNA
is represented by LDOC1S?
22. CDS!5’UTR! 3’UTR! CDS!5’UTR! 3’UTR!
Common
TaqMan Assay
Isoform-Specific
TaqMan Assays
Isoform-Specific TaqMan Assays
Splice Variant transcript
(LDOC1S)
Wild Type LDOC1 transcript
(LDOC1)
Duzkale H. et al., Blood 2011, 117(15):4076-84
23. CDS!5’UTR! 3’UTR! CDS!5’UTR! 3’UTR!
Artificial
Template
LDOC1
Forward
Primer
Reverse
Primer
TaqMan Probe
(Taq-LDOC1) Artificial
Template
LDOC1S
Forward
Primer
Reverse
Primer
TaqMan Probe
(Taq-LDOC1S)
Isoform-Specific TaqMan Assays
Splice Variant transcript
(LDOC1S)
Wild Type LDOC1 transcript
(LDOC1)
Duzkale H. et al., Blood 2011, 117(15):4076-84
Common
TaqMan Assay
Isoform-Specific
TaqMan Assays
24. DCt TaqLD1sv (Ct target LD1svTemplate-Ct homolog LD1wtTemplate)]= 2^-(11.012-26.415) = 43,327.6
DCt TaqLD1wt (Ct target LD1wtTemplate-Ct homolog LD1svTemplate)]= 2^-(10.358-34.104) = 14,068,839.6
Taq-LDOC1S/LDOC1S Taq-LDOC1S/LDOC1
Taq-LDOC1/LDOC1 Taq-LDOC1/LDOC1S
Water/Taq-LDOC1
24 cycles
14.5 cycles
TaqMan Assays
Highly Specific
for
Their Target
Isoforms
Duzkale H. et al., Blood 2011, 117(15):4076-84
25. LDOC1 Isoforms are Differentially Expressed in
Normal and Malignant B cells and Solid Tumor Cell Lines
[Error bars: SE of ΔΔ Ct values]
Duzkale H. et al., Blood 2011, 117(15):4076-84
RelativeLevelsofmRNA
RelativemRNAExpression
26. LDOC1 Isoforms are Differentially Expressed in
Normal and Malignant B cells and Solid Tumor Cell Lines
-LDOC1S mRNA constitutes a small portion of total LDOC1 mRNA
-Potential role for LDOC1 in B cell development, activation,
differentiation and malignant transformation…
[Error bars: SE of ΔΔ Ct values] Duzkale H. et al., Blood 2011, 117(15):4076-84
RelativemRNAExpression
28. Functional studies
LDOC1 protein
knock-down in
HeLa cells
Transcriptome
Biological
Outcome
Proliferation?
Cell cycle?
Network
Construction
(Ingenuity Pathways)
Assessment in HeLa
(QRT-PCR)
STRATEGY
Validation in CLL samples
(MF-QRT-PCR)
Concordant Target
Gene Assessment
29. Transient Transfection of HeLa Cells
with siRNAs
Untransfected
LDOC1
siRNA Pool
Non-targeting
siRNA Pool
Mock
Transfection
siGlo
Cell number, viability & cell cycle
Transfection Efficiency (IF, WB)
RNA
3 biologic replicates/each
34. Global Gene Expression Profiling
of HeLa Cells
Affymetrix Human Genome Chips U133 Plus 2.0
47,000 transcripts with 54,675 probe sets/array
LDOC1 siRNA pool NonTargeting siRNA pool
Biologic replicates Biologic replicates
36. Networks by Ingenuity Pathways Analysis
1. Cardiac Arrythmia,Cardiovascular Disease, Gene Expression
2. Cellular Function and Maintenance, Cancer, GI Disease
3. Cell Cycle, Expression, Cellular Growth and Proliferation
4. Cellular assembly and Organization, Cardiac Necrosis/Cell death, Cell Death
5. Gene Expression, Cell Death, Tissue Development
6. DNA Replication, Recombination and Repair, Cardiovascular Disease, Cancer
38. HeLa cells
(LDOC1 or )
30 CLL samples
(LDOC1 or )
Concordant Target Gene Search-
Intersection of Affymetrix Data
Intersection
Affymetrix Gene Chip
Human Genome U133 Plus 2.0
47,000 transcripts
Affymetrix Gene Chip
Human Genome U133A 2.0
18,400 transcripts
(Abruzzo LV, et al., J Mol Diagn. 2005 7(3):337-45)
39. Definition of LDOC1 Positivity in
Affymetrix Data, Aided by QRT-PCR Data
LDOC1mRNAExpression(QRT-PCR)
LDOC1 mRNA Expression (Affymetrix arrays)
40. Concordant Target Genes in HeLa and CLL samples-
Intersection of Affymetrix Data
(N=51; p<0.05)
CLL Fold Change = LDOC1 high CLL / LDOC1 low CLL
HeLa Fold Change = LDOC1 high (NT siRNAs) / LDOC1 low (LDOC1 siRNAs)
41. Concordant Target Genes in HeLa and CLL samples-
Validation by QRT-PCR in HeLa
ITGAL (Integrin alpha-L)
MAPKAPK2 (MAPK-activated protein kinase 2)
CLCN4 (chloride channel 4)
DNAJA1 (DnaJ homolog subfamily A member 1)
UBE2N (E2 ubiquitin conjugating enzyme)
NRIP1 (nuclear receptor interaction protein)
LDOC1
(3 biologic replicates, triplicate/each)
42. Except for LDOC1, none of the genes
was differentially expressed in HeLa cells
by QRT-PCR assay…
43. Abruzzo L.V. et al., submitted
Screening of CLL literature for candidate
biomarkers of prognosis in CLL
Evaluation of 43 genes in
76 untreated CLL patients
(MF-QRT-PCR)
30 genes were validated as
differentially expressed between
LDOC1 CLL and LDOC1 CLL
Validation of Biomarkers of Prognosis in CLL
with respect to LDOC1 Status
44. Assessment of 43 biomarkers of CLL prognosis
in HeLa cells (LDOC1 or )
Concordant Target Gene Identification
(MF-QRT-PCR)
45. Expression of 43 Biomarkers of CLL Prognosis
in HeLa Cells (by MF-QRT-PCR)
46. Validation in
HeLa cells
LDOC1 or
[3 genes]
Validation in
76 CLL samples
LDOC1 or
[30 genes]
Concordant Target Gene Identification
(MF-QRT-PCR)
Growth Factor-Independent 1
(GFI1)
47. Growth Factor-Independent 1
(GFI1)
-zinc finger transcriptional repressor
-confers IL-2-independent proliferation to
IL-2-dependent T cell lymphoma cell line
-enhances proliferation and inhibits apoptosis
-targets: p21 & p15 (indirect), Bax (direct)
-induces accelerated lymphoma in combination
with Pim-1 or L-myc in mice
-restricts hematopoetic stem cell proliferation;
preserves self-renewal
-expressed highest in early B- and T-cells; low/absent
in mature lymphocytes; increases by TCR stimulation
Zinc finger 268;
by
Thomas
Splettstoesser
49. Bax
GFI1 Inhibits Apoptosis by Repressing
Transcription of Pro-apoptotic Bax
GFI1
based on the data from:
Grimes H.L. et al., 1996 PNAS 93: 14569-14573
ApoptosisX
50. CDKN1A (p21)
CDKN2B (p15)
Miz1 Induces Transcription of CDKN1A and CDKN2B
and Causes Cell Cycle Arrest
Miz1
adapted from:
Liu Q. et al., 2010 Oncogene 29: 2843-2852 &
Basu S. et al., 2009 PNAS 106(5): 1433-1438
Cell cycle arrest
51. GFI1
Miz1
c-Myc
GFI1 and c-Myc Promote Cell Proliferation by
Repressing Transcription of CDKN1A and CDKN2B
through Miz1
CDKN1A (p21)
CDKN2B (p15)
adapted from:
Liu Q. et al., 2010 Oncogene 29: 2843-2852 &
Basu S. et al., 2009 PNAS 106(5): 1433-1438
Cell cycle arrestX
52. LDOC1 may Directly Regulate GFI1 Transcription
to Inhibit Apoptosis and Cell Cycle Arrest
Apoptosis
Cell cycle arrest
GFI1
-In HeLa cells high LDOC1 expression is associated with
high GFI1 mRNA, low Bax and CDKN1A mRNA expression
-LDOC1 & GFI1 are highly expressed in aggressive CLL
LDOC1
Bax
CDKN1A (p21)
X
53. GFI1
Miz1
c-Myc
LDOC1 might Indirectly Interact with GFI1
to Promote Proliferation
CDKN1A (p21)
LDOC1
-In HeLa cells high LDOC1 expression is associated with
high GFI1 mRNA and low CDKN1A mRNA expression
Cell cycle arrestX
-LDOC1 & GFI1 are highly expressed in aggressive CLL
54. CONCLUSIONS
1. LDOC1 mRNA is a novel biomarker of survival
in untreated CLL patients
2. LDOC1S is a new splice variant of LDOC1
3. LDOC1 knock-down in HeLa cells perturbs
genes expressed in cancer-related networks
4. Interaction of LDOC1 and GFI1 may contribute
to the aggressive behavior in a subset of CLL
55. FUTURE DIRECTIONS
1. Evaluate LDOC1 mRNA as a biomarker in a
larger CLL patient cohort in a longitudinal study
2. Elucidate LDOC1-GFI1 interaction
3. Investigate dynamic interplay between
GFI1 and LDOC1 in a system relevant to
CLL pathogenesis
-in vitro stimulation of CLL samples
-BCR, CD40L/IL4,TLR…PI3K/Akt pathway
56. Acknowledgments
Abruzzo Lab
Lynne V. Abruzzo, MD, PhD
Lynn L. Barron, BS
Katherine Lin, MD
Roberto Nussenzveig, PhD
Carmen D. Schweighofer,
MD Wilkinson Lab
Wai Kin Chan, PhD
Lulu Huang, PhD
Hematopathology
Kaushali Patel, MS
Marco Herling, MD
Elias Drakos, PhD
Lan Pham, PhD
Evan Cohen, BS
Committee Members
Shelly Barton, PhD
Emil J Freireich, MD, DSci (Hon)
Vicki Huff, PhD
Kevin R. Coombes, PhD
David McConkey, PhD
Richard Ford, MD, PhD
Malcolm Brenner, MD, PhD
Gil Cote, PhD
Craig Logsdon, PhD
UT GSBS
George Stancel, PhD
Jon R. Wiener, PhD
U.T. Center for Clinical and
Translational Sciences (CCTS)
NIH T32 Training Award