Development of iPS Cells for Treatment of Hemoglobinopathies

Summary

Principal Investigator: Yuet Wai Kan
Abstract: DESCRIPTION (provided by applicant): The proposed POI is a multi-institutional grant that will develop a stem cell based therapy for the treatment of sickle cell disease (SCD) and [unreadable]-thalassemia ([unreadable]-thal) as well as other hemoglobinopathies, using patient derived somatic cells and reprogramming them into induced pluripotent stem (IPS) cells that will have their mutations corrected and ultimately differentiated into hematopoietic stem cells (HSCs) to reconstitute the patient's hematopoietic system. Development of an effective cellular therapy for the treatment of hemoglobinopathies, the most common inherited diseases worldwide, would significantly improve the quality of life of individuals afflicted with SCD and B-thalassemia that are common among the peoples of Africa, the Mediterranean, the Middle East, and Asia as well as their descendents in the U.S. This proposal will test the hypothesis that an effective cellular and genetic therapy for these diseases can be achieved in the context of this PPG through the generation, modification, and the hematopoietic differentiation of patient derived iPS cells. This will be accomplished through the following Projects: Project 1 will involve the conversion of a patient's somatic cells into IPS cells using phiC31 Integrase-mediated, sequence-specific integration of a plasmid carrying 2A peptide linked Oct4, Sox2, Klf4, and cMyc reprogramming cDNAs or by using small activating double stranded RNA (saRNA) to transiently enhance the expression of these reprogramming genes. Project 2 will involve correction of the disease causing mutations in the somatic cells and the iPS cells by sequence specific modification using either classical homologous recombination (HR) or by oligo/polynucleotide-based small fragment homologous replacement (SFHR) in the presence or absence of targeted zinc finger nucleases (ZFNs) or other meganucleases. Project 3 will involve exposure of uncorrected and corrected iPS cells to conditions to direct hematopoietic differentiation to generate HSCs which have the capacity to engraft and reconstitute the hematopoietic system. In the course of this PPG, all Projects will develop xeno-free systems to optimize safety. The science in the Projects will be augmented by an administrative (Core A) and 2 scientific Cores: Core B: Cell and Molecular Biology, and Core C: Cell Transplantation and Analysis.
Funding Period: 2011-09-30 - 2016-07-31
more information: NIH RePORT

Top Publications

  1. pmc A global DNA methylation and gene expression analysis of early human B-cell development reveals a demethylation signature and transcription factor network
    Seung Tae Lee
    Department of Epidemiology and Biostatistics, University of California San Francisco, San Francisco, CA 94158, USA
    Nucleic Acids Res 40:11339-51. 2012
  2. pmc Identification of an astrovirus commonly infecting laboratory mice in the US and Japan
    Terry Fei Fan Ng
    Blood Systems Research Institute, San Francisco, California, United States of America
    PLoS ONE 8:e66937. 2013
  3. pmc Blood cell-derived induced pluripotent stem cells free of reprogramming factors generated by Sendai viral vectors
    Lin Ye
    Department of Medicine and Institute for Human Genetics, University of California, San Francisco, San Francisco, CA 94143, USA
    Stem Cells Transl Med 2:558-66. 2013
  4. pmc Production of factor VIII by human liver sinusoidal endothelial cells transplanted in immunodeficient uPA mice
    Marina E Fomin
    Blood Systems Research Institute, San Francisco, California, United States of America Department of Laboratory Medicine, University of California San Francisco, San Francisco, California, United States of America
    PLoS ONE 8:e77255. 2013
  5. pmc Nuclease-mediated double-strand break (DSB) enhancement of small fragment homologous recombination (SFHR) gene modification in human-induced pluripotent stem cells (hiPSCs)
    R Geoffrey Sargent
    Department of Otolaryngology Head and Neck Surgery, University of California, San Francisco, San Francisco, CA, USA
    Methods Mol Biol 1114:279-90. 2014
  6. pmc The adult livers of immunodeficient mice support human hematopoiesis: evidence for a hepatic mast cell population that develops early in human ontogeny
    Marcus O Muench
    Blood Systems Research Institute, San Francisco, California, United States of America Laboratory Medicine, University of California San Francisco, San Francisco, California, United States of America Liver Center, University of California San Francisco, San Francisco, California, United States of America
    PLoS ONE 9:e97312. 2014

Research Grants

  1. Novel Methods to Assess the Effects of Chemicals on Child Development
    Susan L Schantz; Fiscal Year: 2013
  2. Endothelial Cell Phenotypes in Health and Disease
    William C Aird; Fiscal Year: 2013
  3. Pathobiology of the Enteric System
    Joseph H Szurszewski; Fiscal Year: 2013
  4. Integrative Metabolic Adaptations to Enviromental and Nutritional Challenge
    MORRIS JAY BIRNBAUM; Fiscal Year: 2013
  5. VASCULAR RELATIONS OF BLOOD CELLS AND PROTEINS
    Richard E Waugh; Fiscal Year: 2013
  6. Thrombus Formation and Antithrombotic Intervention
    John H Griffin; Fiscal Year: 2013
  7. INSULIN RECEPTORS AND THE GLUCOSE TRANSPORT SYSTEM
    JERROLD MICHAEL OLEFSKY; Fiscal Year: 2013
  8. Immune Cells in Atherosclerosis and Vascular Disease
    Catherine C Hedrick; Fiscal Year: 2013
  9. Cardiac Fibrillation: Mechanisms and Therapy
    James N Weiss; Fiscal Year: 2013
  10. DEVELOPMENTAL BIOLOGY OF HUMAN ERYTHROPOIESIS
    Stuart H Orkin; Fiscal Year: 2013

Detail Information

Publications7

  1. pmc A global DNA methylation and gene expression analysis of early human B-cell development reveals a demethylation signature and transcription factor network
    Seung Tae Lee
    Department of Epidemiology and Biostatistics, University of California San Francisco, San Francisco, CA 94158, USA
    Nucleic Acids Res 40:11339-51. 2012
    ....
  2. pmc Identification of an astrovirus commonly infecting laboratory mice in the US and Japan
    Terry Fei Fan Ng
    Blood Systems Research Institute, San Francisco, California, United States of America
    PLoS ONE 8:e66937. 2013
    ..This study demonstrates the need for metagenomic screening of laboratory animals to identify adventitious infections that may affect experimental outcomes. ..
  3. pmc Blood cell-derived induced pluripotent stem cells free of reprogramming factors generated by Sendai viral vectors
    Lin Ye
    Department of Medicine and Institute for Human Genetics, University of California, San Francisco, San Francisco, CA 94143, USA
    Stem Cells Transl Med 2:558-66. 2013
    ..Maintenance of the genomic integrity of iPSCs without integration of exogenous DNA should allow the development of therapeutic-grade stem cells for regenerative medicine...
  4. pmc Production of factor VIII by human liver sinusoidal endothelial cells transplanted in immunodeficient uPA mice
    Marina E Fomin
    Blood Systems Research Institute, San Francisco, California, United States of America Department of Laboratory Medicine, University of California San Francisco, San Francisco, California, United States of America
    PLoS ONE 8:e77255. 2013
    ..Demonstration of human FVIII production by transplanted LSECs encourages further pursuit of LSEC transplantation as a cellular therapy for the treatment of hemophilia A. ..
  5. pmc Nuclease-mediated double-strand break (DSB) enhancement of small fragment homologous recombination (SFHR) gene modification in human-induced pluripotent stem cells (hiPSCs)
    R Geoffrey Sargent
    Department of Otolaryngology Head and Neck Surgery, University of California, San Francisco, San Francisco, CA, USA
    Methods Mol Biol 1114:279-90. 2014
    ..Using an allele-specific PCR (AS-PCR)-based cyclic enrichment protocol, clonal populations of corrected CF-iPS cells were isolated and expanded. ..
  6. pmc The adult livers of immunodeficient mice support human hematopoiesis: evidence for a hepatic mast cell population that develops early in human ontogeny
    Marcus O Muench
    Blood Systems Research Institute, San Francisco, California, United States of America Laboratory Medicine, University of California San Francisco, San Francisco, California, United States of America Liver Center, University of California San Francisco, San Francisco, California, United States of America
    PLoS ONE 9:e97312. 2014
    ..Thus, offering a model system to study the interaction of diverse human liver cell types that regulate hematopoiesis and immune function in the liver...

Research Grants62

  1. Novel Methods to Assess the Effects of Chemicals on Child Development
    Susan L Schantz; Fiscal Year: 2013
    ..This Center's research will fill an important gap in our knowledge by investigating the effects of these chemicals, both alone and combination with a high fat diet on reproductive and neural development. ..
  2. Endothelial Cell Phenotypes in Health and Disease
    William C Aird; Fiscal Year: 2013
    ..Core C ("Gene Targeting Core";William C. Aird, Core Leader) provides the necessary tools for targeting the Hprt locus and the loci of endogenous genes in ES cells and mice. ..
  3. Pathobiology of the Enteric System
    Joseph H Szurszewski; Fiscal Year: 2013
    ..This highly-integrated Program will make significant progress toward understanding the pathobiology of the enteric system in gastric emptying disorders and translate this knowledge into new diagnostic tools and therapy. ..
  4. Integrative Metabolic Adaptations to Enviromental and Nutritional Challenge
    MORRIS JAY BIRNBAUM; Fiscal Year: 2013
    ..The projects are supported by three Cores that provide histochemical analysis, generation of genetically modified mice, and their metabolic phenotyping. ..
  5. VASCULAR RELATIONS OF BLOOD CELLS AND PROTEINS
    Richard E Waugh; Fiscal Year: 2013
    ..The underlying mechanisms for these involve mechanical forces, molecular interactions and cellular properties acting synergistically in ways that are uniquely addressed by this program. ..
  6. Thrombus Formation and Antithrombotic Intervention
    John H Griffin; Fiscal Year: 2013
    ..New knowledge will contribute to improving prevention, diagnosis and treatment of relevant diseases related to thrombosis. ..
  7. INSULIN RECEPTORS AND THE GLUCOSE TRANSPORT SYSTEM
    JERROLD MICHAEL OLEFSKY; Fiscal Year: 2013
    ..These studies should lead to an improved understanding of the basic causes of Type 2 diabetes mellitus and hold the potential for new therapeutic modalities. ..
  8. Immune Cells in Atherosclerosis and Vascular Disease
    Catherine C Hedrick; Fiscal Year: 2013
    ..Each project will utilize the Human Core to study immune cells from human subjects to establish functional links between candidate genes of interest and immune cell function in atherogenesis. ..
  9. Cardiac Fibrillation: Mechanisms and Therapy
    James N Weiss; Fiscal Year: 2013
    ..Together, these studies will provide critical groundwork necessary to develop and advance novel therapies for this major complication and cause of mortality from heart disease. ..
  10. DEVELOPMENTAL BIOLOGY OF HUMAN ERYTHROPOIESIS
    Stuart H Orkin; Fiscal Year: 2013
    ..abstract_text> ..
  11. Program Project: Growth, Differentiation and Disease of Urothelium
    Tung Tien Sun; Fiscal Year: 2013
    ..abstract_text> ..
  12. PPG - Gene Therapy for Cystic Fibrosis Lung Disease
    Paul B McCray; Fiscal Year: 2013
    ..The discoveries from this PPG will accelerate the development of gene-based medicine for patients who suffer from this devastating disease...
  13. The Virtual Physiological Rat Project
    Daniel A Beard; Fiscal Year: 2013
    ..This proposal targets the grand challenge of understanding complex multi-faceted disease phenotypes through experiments and simulations that capture the complex genotype-environment-phenotype relationship. ..
  14. Basic and Clinical Studies of Cystic Fibrosis
    Raymond A Frizzell; Fiscal Year: 2013
    ..The Core Center will operate a Pilot and Feasibility Program to bring new investigators into CF research. This Center emphasizes the translation of basic knowledge into applied therapeutics. ..
  15. UNMC EPPLEY CANCER CENTER SUPPORT GRANT
    Kenneth H Cowan; Fiscal Year: 2013
    ....
  16. Role of Eosinophils in Airway Inflammation and Remodeling
    Nizar N Jarjour; Fiscal Year: 2013
    ..Given the prominence of eosinophilic inflammation in a significant proportion of severe asthma patients, these advances will have direct implications for the patients most affected by this very common illness. ..
  17. Inflammatory responses of vascular cells
    Paul L Fox; Fiscal Year: 2013
    ..abstract_text> ..
  18. HORMONAL REGULATION OF BLOOD PRESSURE
    Michal Laniado Schwartzman; Fiscal Year: 2013
    ..ular tone, in the pathophysiology of hypertension and cardiovascular disease. ..
  19. Digitalis-Induced Signaling by Cardiac Na+/K+-ATPase
    Amir Askari; Fiscal Year: 2013
    ..abstract_text> ..
  20. EARLY EVENTS IN ALZHEIMER PATHOGENESIS
    SUE TILTON GRIFFIN; Fiscal Year: 2013
    ..The synergy between our aims, approaches, and measures will enable us to meet our goal of defining early cellular interactions toward development of rational interventions in AD. ..
  21. DIABETES AND ENDOCRINOLOGY RESEARCH CENTER
    Domenico Accili; Fiscal Year: 2013
    ....
  22. Expanding Excellence in Developmental Biology in Oklahoma
    Linda F Thompson; Fiscal Year: 2013
    ..abstract_text> ..
  23. Jules Stein Eye Institute Core Grant for Vision Research
    Wayne L Hubbell; Fiscal Year: 2013
    ..Support in the form of the Core grant is requested to maintain these Modules through instrument service contracts, and to provide necessary personnel support to assist and train users and provide routine maintenance. ..
  24. Functional Consequences of Impaired Autophagy in Aging
    ANA M CUERVO; Fiscal Year: 2013
    ..Significance: These studies may ultimately lead to fundamental insights for understanding, treating or preventing the metabolic alterations and declined cognitive and immune function characteristic of elders. ..
  25. CARDIOVASCULAR DYNAMICS AND THEIR CONTROL
    John E Hall; Fiscal Year: 2013
    ..End of Abstract) ..
  26. The Center for Native and Pacific Health Disparities Research
    MARJORIE K LEIMOMI MALA MAU; Fiscal Year: 2013
    ..5) To prepare and empower our diverse Native and Pacific People communities to take ownership of their own health and wellness. ..
  27. Molecular and Cellular Basis for Digestive Diseases
    Richard M Peek; Fiscal Year: 2013
    ..The Administrative Core also contains Biostatistics and Enrichment Programs and oversees the financial management and operation of the VDDRC. ..
  28. Neurohumoral control of veins in hypertension
    Gregory D Fink; Fiscal Year: 2013
    ..This project tests the idea that altered structure or function of veins also may cause hypertension, and that it may be possible to treat hypertension using drugs that affect veins. ..
  29. Rocky Mountain Regional Center of Excellence or Biodefense and Emerging Infectiou
    John T Belisle; Fiscal Year: 2013
    ..abstract_text> ..
  30. Regulatory Mechanisms In Intestinal Motility
    Kenton M Sanders; Fiscal Year: 2013
    ..The investigative team is highly synergistic and collaborative, and the PPG has a long track-record of productivity and novel discovery ..
  31. Mechanistic Pharmacology of Anti-Mitotics and Apoptosis Regulation
    Timothy J Mitchison; Fiscal Year: 2013
    ..In aim 4 we will pursue several approaches towards translating mechanistic understanding from aims 1-3 into improved patient care. ..
  32. Osteocyte Regulation of Bone/Muscle with Age
    Lynda F Bonewald; Fiscal Year: 2013
    ..The results of these experiments should lead to novel therapeutics for the prevention and treatment of both osteoporosis and sarcopenia. ..
  33. Gene Networks controlling macrophage-adipocyte interactions in insulin
    Christopher K Glass; Fiscal Year: 2013
    ..abstract_text> ..
  34. STEM CELL GENE THERAPY FOR HEMOGLOBINOPATHIES
    George Stamatoyannopoulos; Fiscal Year: 2013
    ..The focus of this Program Project, Gene Therapy, can provide a new paradigm for the treatment of these hemoglobinopathies as well as for other blood diseases. (End of Abstract) ..
  35. Metabolic effects of adipose lipogenesis
    Timothy E McGraw; Fiscal Year: 2013
    ....
  36. Invertebrate Models of Fat Storage
    Jonathan M Graff; Fiscal Year: 2013
    ..In Aim III, we will characterize the role that this E3 ligase has in fat biology;knowledge that is key in order to develop the fundamental insights required to ultimately manipulate the Adp pathway for therapeutic ends. ..
  37. The Role of H6PDH and 11beta-HSD1 in Type 2 Diabetes and Obesity
    Yanjun Liu; Fiscal Year: 2013
    ..Blocking the effects of H6PDH on 112-HSD1 may represent a new strategy in the effective treatment of type 2 diabetes and obesity through reduction of tissue GC availability mediating insulin sensitivity and glucose homeostasis. ..
  38. PAHs: New Technologies and Emerging Health Risks
    David E Williams; Fiscal Year: 2013
    ..Accomplishing these goals will provide significant scientific advancement and improve the quality of life for impacted communities. ..