Lab Research Training In Pediatric Oncology-Hematology
Curt I Civin, Herman & Walter Samuelson Professor Of C
Oncology Centerjohns Hopkins University
Grant 2T32CA060441-16 from National Cancer Institute, IRG: NCI
Abstract: Program in Laboratory Research in Pediatric Oncology-Hematology To continue to make progress in Pediatric Oncology-Hematology (POH), we need to perpetuate the pool of talented and trained POH physician-scientists and PhD scientists who will make POH-relevant discoveries and provide investigative leadership. Nevertheless, in the field of POH, there continues to be a shortage of young investigators with extensive laboratory research expertise. In the current funding period (2002-2008), this grant has already supported superb laboratory research training for an outstanding group of 21 physician-scientists and 8 PhD scientists who have demonstrated a high degree of productivity and commitment to careers in cancer research. In addition, these trainees have strengthened the laboratory research base in POH in the Johns Hopkins Medical Institutions (JHMI). This application again seeks support for 10 interdisciplinary postdoctoral research fellowship positions per year to fund the research training that is the major ingredient of the recently combined Johns Hopkins University/National Cancer Institute (NCI) Subspecialty Fellowship Training Program in POH (Joint Fellowship Training Program). Two groups of postdoctoral fellows will be supported by the grant proposed herein. The major group of trainees will be composed of physician-scientists, since the laboratory research training of academic POH physician-scientists is the primary objective of this training program. These highly selected physician- scientists will have completed the clinical year of their fellowships in the Joint Fellowship Training Program. The second (smaller) group of trainees will be composed of PhD postdoctoral scientists working at JHMI on projects highly relevant to the biology of childhood cancer and blood diseases. These strongly qualified postdoctoral trainees will be provided intensive 3-year research experiences that will prepare them for careers in cancer research. Trainees will work on projects relevant to the biology of childhood cancer and blood diseases, in the laboratories of leading scientists in the field. The trainees and their mentors will participate in an interactive, dynamic research environment bridging the JHMI and the National Institutes of Health (NIH), with an emphasis on translational research. Summary of relevance to public health This program provides postdoctoral education, training, and experience in field-leading research on childhood cancer and blood diseases, principally for physician- scientist trainees during their subspecialty fellowships in pediatric oncology-hematology
Project start date: 1993-09-30
Project end date: 2013-06-30
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Lab Research Training In Pediatric Oncology-Hematology
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 2T32CA060441-11 from National Cancer Institute, IRG: NCI
Abstract: To continue to make progress in Pediatric Oncology-Hematology (POH), we need to perpetuate the pool of talented and trained POH physician-scientists and Ph.D. scientists who will make POH-relevant discoveries and provide investigative leadership. Nevertheless, in the field of POH, there continues to be a shortage of young investigators with extensive laboratory research expertise. In the current funding period (1998-2003), this grant has already supported superb laboratory research training for an outstanding group of 17 physician-scientists and 12 Ph.D. scientists who have demonstrated a high degree of productivity and commitment to careers in cancer research. Selection of the 2002-2003 trainees is in progress. In addition, these trainees have strengthened the laboratory research base in the POH in the Johns Hopkins Medical Institutions (JHMI). This application now seeks support for 10 interdisciplinary postdoctoral research fellowship positions per year to fund the research training that is the major ingredient of the recently combined Johns Hopkins University/National Cancer Institute (NCI) Subspecialty Fellowship Training Program in POH (JHU/NCISFTP). Two groups of postdoctoral fellows will be supported by the grant proposed herein. The major group of trainees will be composed of physician-scientists, since the laboratory research training of academic POH physician-scientists is the primary objective of this training program. These highly selected physician-scientists will have completed the clinical year of their fellowships in the JHU/NCISFTP. The second (smaller) group of trainees will be composed of Ph.D. postdoctoral scientists working at JHMI on projects highly relevant to the biology of childhood cancer and blood diseases. These strongly qualified postdoctoral trainees will be provided intensive 3-year research experiences that will prepare them for careers in cancer research. Trainees will work on projects relevant to the biology of childhood cancer and blood diseases, in the laboratories of leading scientists in the field. The trainees and their mentors will participate in an interactive, dynamic research environment bridging the JHMI and the National Institutes of Health (NIH), with an emphasis on translational research.
Project start date: 1993-09-30
Project end date: 2008-05-31
2T32CA060441-11 (2003): $547684
2T32CA060441-06 (1998): $276887
Grants awarded to Curt I Civin
Lab Research Training In Pediatric Oncology-Hematology
Curt I Civin, Herman And Walter Samuelson Professor Of C
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 5T32CA060441-15 from National Cancer Institute, IRG: NCI
Abstract: To continue to make progress in Pediatric Oncology-Hematology (POH), we need to perpetuate the pool of talented and trained POH physician-scientists and Ph.D. scientists who will make POH-relevant discoveries and provide investigative leadership. Nevertheless, in the field of POH, there continues to be a shortage of young investigators with extensive laboratory research expertise. In the current funding period (1998-2003), this grant has already supported superb laboratory research training for an outstanding group of 17 physician-scientists and 12 Ph.D. scientists who have demonstrated a high degree of productivity and commitment to careers in cancer research. Selection of the 2002-2003 trainees is in progress. In addition, these trainees have strengthened the laboratory research base in the POH in the Johns Hopkins Medical Institutions (JHMI). This application now seeks support for 10 interdisciplinary postdoctoral research fellowship positions per year to fund the research training that is the major ingredient of the recently combined Johns Hopkins University/National Cancer Institute (NCI) Subspecialty Fellowship Training Program in POH (JHU/NCISFTP). Two groups of postdoctoral fellows will be supported by the grant proposed herein. The major group of trainees will be composed of physician-scientists, since the laboratory research training of academic POH physician-scientists is the primary objective of this training program. These highly selected physician-scientists will have completed the clinical year of their fellowships in the JHU/NCISFTP. The second (smaller) group of trainees will be composed of Ph.D. postdoctoral scientists working at JHMI on projects highly relevant to the biology of childhood cancer and blood diseases. These strongly qualified postdoctoral trainees will be provided intensive 3-year research experiences that will prepare them for careers in cancer research. Trainees will work on projects relevant to the biology of childhood cancer and blood diseases, in the laboratories of leading scientists in the field. The trainees and their mentors will participate in an interactive, dynamic research environment bridging the JHMI and the National Institutes of Health (NIH), with an emphasis on translational research.
Project start date: 1993-09-30
Project end date: 2008-05-31
5T32CA060441-15 (2007): $480645
5T32CA060441-14 (2006): $499219
5T32CA060441-13 (2005): $587013
5T32CA060441-12 (2004): $587714
Hematopoietic Stem Cells For Transplantation
Curt I Civin, Herman And Walter Samuelson Professor Of C
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 2P01CA070970-10A1 from National Cancer Institute, IRG: ZCA1
Abstract: This grant proposal focuses our sustained laboratory and clinical research experience in animal and human stem cell biology and leukemia to extend exciting new investigations on normal tissue-specific and cancer stem cells. The investigators in this PPG look forward to discovering key information about the defining characteristics of cancer stem cells. This knowledge will enable us to develop new methods to better identify and isolate these rare cells for further study. In addition, this PPG seeks increased understanding of the pathophysiologic mechanisms of cancer stem cells, particularly the signaling pathways that these cells depend on for their persistence and other malignant behavior. This basic information will suggest new clinical cancer stem cell targets, which we will proceed to attack via pharmacologic and biologic approaches. Both within the Projects and in interactions among the Projects and Cores, clinical research and laboratory inquiries will proceed in parallel, so that insights generated in one sphere will translate to, and promote progress in the other. We believe that our highly interactive, closely integrated, translational approach is the best way to determine mechanisms of normal and malignant stem cell biology, and simultaneously to develop new treatments utilizing normal stem cells and targeting malignant stem cells in human diseases. Relevance There is growing evidence suggesting that many cancers originate in early (stem or progenitor) cells and that the mature cancers continue to be maintained by a stem-progenitor cell hierarchy. Our group has provided important evidence demonstrating the existence of discrete populations of cancer stem cells in human diseases, and our results support the prediction that cancer recurrence in patients after chemotherapy may be due to failure to eradicate the rare, sustaining cancer stem cell population, despite massive reduction of the predominant bulk population of mature cancer cells. The investigators in this PPG look forward to discovering key information about the defining characteristics and molecular mechanisms of cancer stem cells that will provide new ways to identify and attack cancer stem cells therapeutically.
Project start date: 2008-02-26
Project end date: 2013-01-31
Fas Ligand+ Cells To Reduce Transplant Rejection
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 3R21HL072229-02S1 from National Heart, Lung, And Blood Institute, IRG: ZRG1
Abstract: The purpose of this application for a supplement in response to PAR-03-063 ("NHLBI Competitive Supplements for Human Embryonic Stem Cell Research") is to enable my lab, which has little prior experience working with human embryonic stem cell (HESC) lines, to enhance our existing aims of my ongoing NHLBI grant (R2.1 HL72229 "Fas ligand* blood cells to reduce transplant rejection"), which is now being pursued using primary mouse and human lympho-hematopoietic stem-progenitor cells (HSCs). The overall goal of my ongoing grant is to investigate the ability of lentivirally (LV) transduced HSCs or dendritic cells (DCs) expressing high levels of Fas ligand (FasL) to reduce the response to alloantigen (Ag) and increase engraftment of allogeneic (allo) HSCs. In Aims 1-2 of this proposal, we proposed to investigate this in cellular immunology and transplant experiments, as well as to elucidate the apoptotic pathways in alloimmune cells. Since transduced FasL v HSCs might potentially be toxic in vivo, we proposed to investigate technologies to limit potential FasL toxicity. A novel FasL v cell therapy approach to reduce graft rejection might eventually be used in clinical allo blood and marrow transplantation (BMT). Furthermore, achievement of stable lympho-hematopoietic chimerism would be predicted to generate tolerance for transplanted allo organs, which is explored in Aim 3. HESCs can be manipulated to generate blood cells, and hematopoietic transplants of HESC-derived HSCs have been proposed with proof-of-principle illustrated in mice. Since immunologic rejection is a major barrier to use of allo HESCs (including HESC-derived HSCs) clinically, a next logical step for us is to investigate whether FasL transduced HESC-derived HSCs/DCs might reduce the alloimmune response. This use of HESC lines is clearly within the scope and a logical extension of the goals and objectives of our ongoing grant. Indeed, we did not propose this in the initial application only because we had absolutely no hands-on experience with HESCs at that time, and HESC lines were then extremely difficult to obtain. In this supplement application, the experiments involving HESCs are confined to Specific Aim 2.2 (Hypothesis 2.2 FasL v HSCs and their progeny will delete attacking alloreactive effector T cells -- extended in this supplement to investigate HESCs-derived "HSCs").
Project start date: 2002-09-30
Project end date: 2005-08-31
3R21HL072229-02S1 (2003): $81750
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 3P01CA070970-04S19001 from National Cancer Institute, IRG:
Abstract: The purpose of Core Component A, the Administrative, Biostatistics, and FACS Core, led by the Program Project PI, Dr. C. Civin, is to provide all 4 projects with (1) central leadership, administrative management, and mentoring; (2) expertise and resources for biostatistical planning and analysis of laboratory and clinical experiments; and (3) expertise and resources for Fluorescence Activated Cell Sorter (FACS) analysis and cell sorting. He will directly supervise the Administrative, Biostatistics, and FACS resources of Core Component A. The Administrative resource will perform the Program Project s administrative management, including budget management, record-keeping and payments for donations of normal bone marrow, log-books and allocations of use and payments for the FACS resource and animals/animal transplant services, and generation of progress reports and related correspondence, maintenance of our weekly seminar series, leadership of working meetings of Program Project and Core Component Leaders and Investigators, and scheduling of regular meetings of our Internal and External Advisory Groups and consideration/execution of their recommendations. The Biostatistics resource will provide biostatistical support to the entire Program Project. The FACS resource will interact with Program Project investigators on all flow cytometry experiments.
Keywords: biomedical facility, flow cytometry, statistics /biometry
Fas Ligand+ Blood Cells To Reduce Transplant Rejection
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 5R21HL072229-03 from National Heart, Lung, And Blood Institute, IRG: ZHL1
Abstract: The ability to stably transduce lymphohematopoietic stem-progenitor cells (HSCs) and progeny cells allows us to genetically engineer HSCs and progeny, especially dendritic cells (DCs), to serve as cellular tools to improve transplantation. Allogeneic (allo) blood and marrow transplantation (BMT) will continue to be an important treatment option in patients with many inherited and acquired diseases. Our recent Preliminary Results indicate that ex vivo or in vivo treatment with transduced FasL+ allog mouse DCs or HSCs suppressed the response to that alloantigen and increased engraftment of allo cells, without immunoablation or evident toxicity to the recipient mice. In Aims 1-2 of this proposal, we propose studies to test the concept and elucidate the principal mechanisms by which transduced DCs or HSCs constitutively expressing Fas ligand (FasL) appear to reduce the T (and NK) cells that mediate graft rejection without overwhelming organ toxicity or general immunologic impairment. Aims 1-2 attempt to accelerate and enhance the natural homeostatic role of FasL during the course of the alloimmune response, with the potential outcome that alloimmune T cells may be killed earlier in their activation/expansion, or killed more potently later during activation-induced cell death (AICD), by transduced FasL+ DCs (Aim 1) or HSCs/progeny (Aim 2), respectively. As we investigate this, we will examine the apoptotic pathways in alloimmune cells. In addition, since we expect that transduced FasL+ HSCs might potentially be toxic in vivo, we will in parallel investigate technologies to limit potential FasL toxicity, eg by transducing only a small percent of high quality HSCs, by employing a FasL deletion mutant that cannot be cleaved to release soluble FasL (sFasL), by eliminating the transduced cells themselves or their FasL expression after tolerance to HSCs has been generated, or by using lineage/stage-specific promoters to restrict FasL expression. A novel FasL+ cell therapy approach to reduce graft rejection may eventually be used in clinical allo BMT. Furthermore, achievement of stable lymphohematopoietic chimerism would also be predicted to generate tolerance for transplanted allo organs or pluripotent stem cells. Therefore, in Aim 3, we will assess tolerance to cardiac allografts in hematopoietic macro-chimeras, generated by this FasL strategy, with or without other immunosuppressive methodologies such as co-stimulatory blockade. SPECIFIC AIM 1 Assess whether FasL+ DCs specifically reduce an alloimmune response and enhance engraftment of allo HSCs SPECIFIC AIM 2 Determine whether FasL+ HSCs generate specific tolerance in allo BMT. SPECIFIC AIM 3 Evaluate tolerance to cardiac allografts in hematopoietic chimeras generated using FasL+ DCs/HSCs.
Keywords: CD95 molecule, artificial immunosuppression, dendritic cell, hematopoietic stem cell, histocompatibility, transplant rejection, apoptosis, autoimmunity, blood treatment, bone marrow transplantation, gene therapy, heart transplantation, immune tolerance /unresponsiveness, isoantigen, nonhuman therapy evaluation, stem cell transplantation, transfection, NOD mouse, SCID mouse, clinical research, human subject, laboratory mouse
Project start date: 2002-09-30
Project end date: 2005-08-31
5R21HL072229-03 (2004): $327000
5R21HL072229-02 (2003): $245250
Fas/FasL Mediated Apoptosis In GVHD And Aplastic Anemia
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 2P01CA070970-05A10002 from National Cancer Institute, IRG:
Abstract: The ability to stably transduce lympho-hematopoietic stem-progenitor cells (HSCs) allows us to genetically engineer HSCs and their progeny to serve as improved cellular tools to treat disease and complications. In Specific Aim 1, we propose preclinical studies in mouse and human models to confirm the concept and elucidate the principal cellular and molecular mechanisms by which Fas ligand-transduced (FasL*) dendritic cells (DCs) or HSCs may selectively kill the T and NK cells that mediate GVHD. The results of Aim 1 will also provide modeling information on the application of transduced FasL+ cells to reduce immune attack against HSCs in severe aplastic anemia (SAA) (Specific Aim 3). Since it is expected that transduced FasL+ cells may be toxic in potential future translational in vivo applications, we will investigate technologies to limit potential FasL toxicity, eg by eliminating the transduced cells (or their FasL expression) after tolerance to HSCs has been generated. A novel transduced FasL+ cell therapy approach to reduce effector lymphocytes attacking host cells in GVHD (Aim 1) and SAA (Aim 3) may eventually be used in transplants for SAA, PNH and other diseases, and a potential clinical trial is outlined in Aim 1. In addition, our accompanying mechanistic studies on apoptotic pathways in alloimmune cells will increase information on the fundamentals of the Fas pathway in the effector cells mediating GVHD (and SAA), which in turn, should increase understanding of death pathways in the biology of (a) alloimmune responses, (b) hematologic malignancies that evade immune surveillance, and (c) transplanted organs (or pluripotent stem cells). In Specific Aim 2, Projects 1, 2 and 4 will cooperate to investigate the potential role of the Fas pathway in the pathophysiology of HSCs from SANPNH patients. Specific Aim 1 To engineer FasL+ host DCs or HSCs to selectively kill the cellular effectors of GVHD. Specific Aim 2 To investigate the role of the Fas pathway in the pathogenesis of SAA and PNH. Specific Aim 3 To investigate whether FasL+ HSCs selectively kill autologous anti-HSC CTLs in SAA.
Keywords: CD95 molecule, aplastic anemia, apoptosis, cell population study, dendritic cell, graft versus host disease, hematopoietic stem cell, hematopoietic tissue transplantation, autoimmune disorder, cytotoxic T lymphocyte, gene therapy, genetic transduction, immunopathology, paroxysmal nocturnal hemoglobinuria, NOD mouse, SCID mouse, clinical research, genetic manipulation, human subject, human tissue
Project start date: 2002-09-30
Project end date: 2003-03-31
Lab Research Training In Pediatric Oncology-Hematology
Curt I Civin, Herman & Walter Samuelson Professor Of C
Oncology Centerjohns Hopkins University
Grant 2T32CA060441-16 from National Cancer Institute, IRG: NCI
Abstract: Program in Laboratory Research in Pediatric Oncology-Hematology To continue to make progress in Pediatric Oncology-Hematology (POH), we need to perpetuate the pool of talented and trained POH physician-scientists and PhD scientists who will make POH-relevant discoveries and provide investigative leadership. Nevertheless, in the field of POH, there continues to be a shortage of young investigators with extensive laboratory research expertise. In the current funding period (2002-2008), this grant has already supported superb laboratory research training for an outstanding group of 21 physician-scientists and 8 PhD scientists who have demonstrated a high degree of productivity and commitment to careers in cancer research. In addition, these trainees have strengthened the laboratory research base in POH in the Johns Hopkins Medical Institutions (JHMI). This application again seeks support for 10 interdisciplinary postdoctoral research fellowship positions per year to fund the research training that is the major ingredient of the recently combined Johns Hopkins University/National Cancer Institute (NCI) Subspecialty Fellowship Training Program in POH (Joint Fellowship Training Program). Two groups of postdoctoral fellows will be supported by the grant proposed herein. The major group of trainees will be composed of physician-scientists, since the laboratory research training of academic POH physician-scientists is the primary objective of this training program. These highly selected physician- scientists will have completed the clinical year of their fellowships in the Joint Fellowship Training Program. The second (smaller) group of trainees will be composed of PhD postdoctoral scientists working at JHMI on projects highly relevant to the biology of childhood cancer and blood diseases. These strongly qualified postdoctoral trainees will be provided intensive 3-year research experiences that will prepare them for careers in cancer research. Trainees will work on projects relevant to the biology of childhood cancer and blood diseases, in the laboratories of leading scientists in the field. The trainees and their mentors will participate in an interactive, dynamic research environment bridging the JHMI and the National Institutes of Health (NIH), with an emphasis on translational research. Summary of relevance to public health This program provides postdoctoral education, training, and experience in field-leading research on childhood cancer and blood diseases, principally for physician- scientist trainees during their subspecialty fellowships in pediatric oncology-hematology
Project start date: 1993-09-30
Project end date: 2013-06-30
Hematopoietic Stem Cells For Transplantation
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 5P01CA070970-09 from National Cancer Institute, IRG: NCI
Abstract: Myelo-immunosuppressive therapy followed by blood and marrow transplantation (BMT) offers great promise for cure of most hematologic malignancies, a number of solid cancers, and a variety of fatal nonmalignant disorders. Nevertheless at present, only a minority of patients with diseases potentially curable by BMT are indeed cured, since many are not eligible for BMT and a significant number who are transplanted die of toxicity or disease recurrence. The ability to identify, isolate, and manipulate lympho-hematopoietic stem-progenitor cells (HSCs) will improve both the availability and outcome of BMT. The overall goals of this competitive renewal Program Project proposal are to better understand the pathophysiology of normal HSCs and abnormal HSCs in human diseases. Based on this understanding, we propose to develop specific laboratory methods to make HSCs better transplantation tools, for severe aplastic anemia (SAA), PNH, MDS and leukemia, and for BMT in general. Specifically, we propose in this Program Project to 1. further improve our novel and highly effective immunoablative but non-myeloablative treatment for SAA (Projects 1, 2), 2. develop increased understanding of the pathogenesis of SAA and PNH (Projects 1, 2, 3, 4), 3. investigate the role and regulation of FLT3 in normal and diseased HSCs (Projects 3, 2, 1), and 4. molecularly describe apoptotic pathway status of T/NK cells mediating graft vs. host disease(GVHD) and SAA, as well as HSCs in SAA and PNH; determine if cells transduced to constitutively express the Fas Ligand (FasL) gene functionally protect allogeneic host cells from immune attack in GVHD and autologous HSCs from immune attack in SAA/PNH (Projects 4, 2, 1).
Keywords: aplastic anemia, hematopoietic stem cell, hematopoietic tissue transplantation, clinical research
Project start date: 1998-06-30
Project end date: 2008-01-31
5P01CA070970-09 (2006): $1901671
5P01CA070970-08 (2005): $1576651
5P01CA070970-07 (2004): $1843415
5P01CA070970-06 (2003): $1796901
2P01CA070970-05A1 (2002): $1440616
MicroRNAs Regulating Erythroid Development
Curt I Civin, Herman & Walter Samuelson Professor Of C
Oncologyjohns Hopkins University
Grant 1R01DK080750-01 from National Institute Of Diabetes And Digestive And Kidney Diseases, IRG: HP
Abstract: MicroRNAs bind to sites in target mRNAs´ 3´UTR and (principally) inhibit translation to protein. Since binding of microRNAs does not depend on full complementarity with their target sequences, microRNAs can bind to and block protein translation of many different mRNAs, and thereby serve as powerful regulatory switches. We profiled microRNA expression in hematopoietic stem-progenitor cells (HSPCs) and combined this data with human HSPC mRNA expression and thermodynamic microRNA-mRNA target predictions to propose that certain HSPC-expressed microRNAs (HE-microRNAs) regulate hematopoiesis. On this bioinformatic basis, we formulated a model for microRNA control of hematopoiesis in which many genes specifying hematopoietic differentiation are expressed by early HSPCs, but held in check by HE-microRNAs. For several target mRNAs important in hematopoiesis, we demonstrated experimentally that translation is actually decreased by HE- microRNAs. Mir-155 potently reduced both myelopoiesis and erythropoiesis of normal human HSPCs, and mir-16 selectively inhibited erythropoiesis. These findings supported our hypotheses, based on predicted target mRNAs, that (a) mir-155 may regulate development at approximately the common myeloid precursor (CMP) stage prior to erythroid commitment, while (b) mir-16 may block development at the stage of a bipotent megakaryocyte-erythroid progenitor (MEP). Thus, these 2 microRNAs may serve as tools to dissect the very early events of erythroid development and offer potential to expand these rare progenitors. A more detailed model of microRNA expression and molecular targets affecting erythropoiesis should provide additional molecular tools for studying erythroid development and function. Since the cells that we studied included rare adult stem cells and various stages of progenitors, we propose in Aim 1 to expand our microRNA profiles of HSPCs to more highly purified subsets of primary human and mouse erythroid progenitors. In addition, we will profile microRNA expression during erythroid development from NIH-approved human embryonic stem cells (hESCs). In Aim 2, we will determine if selected individual microRNAs experimentally inhibit development of erythroid progenitors. In Aim 3, we will study the molecular mechanisms of the microRNA´s erythropoietic effects by identifying proteins whose synthesis is inhibited by each functionally-active microRNA. This project will investigate the effects of new regulatory molecules called microRNAs, which appear to play novel, potent roles in control of red blood cell formation. Understanding the actions of these microRNAs may provide new tools, for both expansion and control of red blood cell development
Project start date: 2008-05-01
Project end date: 2011-04-30
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 3P01CA070970-04S10002 from National Cancer Institute, IRG:
Abstract: Lymphohematopoietic stem/progenitor cell biology is the long-term interest of Dr. Civin s laboratory. In addition, as clinical hematologist- oncologists, we desire to make direct application of our work to clinical treatment, especially stem/progenitor cell transplantation. Further understanding of the biology of human stem/progenitor cells is essential for many basic questions with clinical relevance, since these are the cells of origin of severe aplastic anemias (SAA), myelodysplastic syndromes (MDS) and leukemias, and because purified stem/progenitor cells are powerful therapeutic tools, especially (though not exclusively) for hematologic disorders. Development for feasible laboratory assays for human lymphohematopoietic stem cells (HSC) is important for basic understanding of hematopoiesis and for clinical BMT. New assays for HSC must be validated by clinical transplantation of candidate HSC, since human transplantation is currently the only way to definitively prove the presence of human HSC in the graft. Specific Aim 1 of Project 2 will extensively characterize "SCID repopulating cells" (SRC), the normal human hematopoietic cells which engraft in irradiated immunodeficient mice, with respect to immunophenotype. SRC will be compared with normal human and murine HSC. Specific Aim 2 will determine the effects of short-term ex vivo culture, an important approach in hematopoietic transplantation, on SRC. Specific Aim 3 will assess the use of SRC assays in cases of SAA, MDS, leukemias, and pediatric solid tumors. This may allow better definition of cancer stem cells, the cells which cause relapse. The laboratory investigation of normal and malignant stem/progenitor cell biology in Project 2 is central to the entire Program Project.
Keywords: cell population study, disease /disorder model, hematopoiesis, hematopoietic stem cell, hematopoietic tissue transplantation, nonhuman therapy evaluation, CD34 molecule, acute lymphocytic leukemia, acute myelogenous leukemia, aplastic anemia, biomarker, dyserythropoietic anemia, embryo /fetus surgery, heterologous transplantation, minimal residual disease, neoplasm /cancer relapse /recurrence, paroxysmal nocturnal hemoglobinuria, pediatric neoplasm /cancer, prognosis, NOD mouse, SCID mouse, clinical research, human fetus tissue, human pregnant subject, human subject
HEMATOPOIETIC STEM CELLS FOR TRANSPLANTATION
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 3P01CA070970-04S1 from National Cancer Institute, IRG: NCI
Abstract: Bone marrow transplantation (BMT) offers great promise for cure of most hematologic malignancies, a number of solid cancers, and a variety of fatal non-malignant disorders. Nevertheless, only a minority of patients with diseases potentially curable by BMT are indeed cured, since many are not eligible for BMT a significant number who are transplanted die of toxicity or disease recurrence. The ability to isolate and expand lymphohematopoietic stem cells (HSC) will improve both the availability and outcome of BMT. Use of purified HSC may avoid histocompatibility problems in allogeneic BMT, as well as improve autologous BMT, for cancer and certain other disease, and for gene therapy. The overall goal of this Program Project (C. Civin, PI) is to study normal HSC clinically and their best surrogates in the laboratory, and to utilize HSC in human transplantation. Clinical and laboratory studies will proceed in parallel; insights generated in one sphere will translate to the other. Project 1 (Jones) will conduct pre-clinical experiments and clinical trials of stem-progenitor cell transplantation for patients with myelodysplastic syndromes (MDS), pediatric solid tumors, severe aplastic anemia (SAA), paroxysmal nocturnal hemoglobinuria (PNH), and autoimmune diseases. In utero transplantation will be investigated for patients with inherited diseases. Project 2 (Civin) will use the human-immunodeficient mouse assay for stem/progenitor cells to answer questions arising from the above clinical trials, to assess purification of candidate human HSC, to test the ex vivo expansion of the engrafting capacity of cord blood stem/progenitor cells, and to measure normal engrafting cells in SAA and cancer stem cells in PNH, MDS, acute leukemias and pediatric solid tumors. Project 3 (Sharkis) will study the key hematopoietic marker, CD34. CD34 null and knock-out mice will be used to determine whether CD34 gene function is required for stem cell self-renewal, proliferation, and/or differentiation. Simultaneously, Project 3 will determine the molecular components of CD34 signal transduction. Project 4 (Small) will concentrate on the FLT3/STK1 receptor. Cis-and trans-regulatory factors important to its expression in normal hematopoiesis and leukemia will be studied. The potential functional role of the FLT3/STK1 molecule in leukemia will be modeled with a constitutively activated receptor.
Keywords: bone marrow transplantation, hematopoiesis, hematopoietic stem cell
Project start date: 1998-06-30
Project end date: 2002-07-31
3P01CA070970-04S1 (2002): $327000
5P01CA070970-04 (2001): $1565574
5P01CA070970-03 (2000): $1502960
5P01CA070970-02 (1999): $1465726
1P01CA070970-01A1 (1998): $1314187
3P01CA070970-09S1 (2007): $775678
3P01CA070970-08S1 (2005): $320731
Lab Research Training In Pediatric Oncology-Hematology
Curt I Civin, Professor Of Oncology And Pediatrics
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 3T32CA060441-13S1 from National Cancer Institute, IRG: NCI
Project start date: 1993-09-30
Project end date: 2008-05-31
3T32CA060441-13S1 (2005): $116780
3T32CA060441-12S1 (2004): $51902
3T32CA060441-10S1 (2002): $39861
5T32CA060441-10 (2002): $399866
5T32CA060441-09 (2001): $327841
5T32CA060441-08 (2000): $264036
5T32CA060441-07 (1999): $339847
5T32CA060441-05 (1997): $135377
5T32CA060441-04 (1996): $127151
5T32CA060441-03 (1995): $179982
5T32CA060441-02 (1994): $184194
1T32CA060441-01 (1993): $142560
DEVELOPMENTAL PROJECT--IMMUNOLOGICAL RESPONSE TO TUMOR ANTIGENS
Curt I Civin
Johns Hopkins University W400 Wyman Park Building Baltimore, Md 212182680
Grant 5P30CA006973-200037 from National Cancer Institute, IRG:
Keywords: NEOPLASMS IMMUNOLOGY, TUMOR ANTIGENS, NEOPLASMS OF BLOOD AND RE SYSTEM, LEUKEMIA ACUTE, NEOPLASMS OF BLOOD AND RE SYSTEM, LEUKEMIA MYELOID, CELL HYBRIDS, HYBRIDOMAS, IMMUNOLOGICAL PREPARATIONS, MONOCLONAL ANTIBODIES, NEOPLASMS, PEDIATRIC, CELL SORTING, LASER, HUMAN, CLINICAL
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