Targeted non-genotoxic hematopoietic stem cell transplant conditioning approach

靶向非基因毒性造血干细胞移植调理方法

• 批准号: 10242836
• 负责人: Shanmuganathan Chandrakasan
• 金额: $ 16.31万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242836
• 关键词: Acute; Address; Adjuvant; Affect; Aging; Alkylating Agents; Animal Model; Animals; Antibodies; Antibody Therapy; Autologous; Award; Bioinformatics; Bone Marrow; Busulfan; CD34 gene; Cell Compartmentation; Cell Cycle; Cell Death; Cell Surface Receptors; Cell Therapy; Cell-Mediated Cytolysis; Cells; Cellular biology; Clinical; Clinical Trials; Coculture Techniques; Complication; Data; Development; Disease; Disease Management; Donor person; Dose; Engraftment; Enhancers; FANCD2 protein; Fanconi Anemia Complementation Group A Protein; Fluorescent Dyes; Funding; Goals; Grant; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hemophilia A; Human; Immune; Immune System Diseases; Immunocompetent; Immunology; Immunotoxins; Infertility; Inherited; Knowledge; Leadership; MPL gene; Malignant - descriptor; Mediating; Mentors; Mentorship; Methods; Modeling; Molecular; Morbidity - disease rate; Mus; Mutagens; Neuraxis; Non-Malignant; Organ; PTPRC gene; Pharmaceutical Preparations; Pharmacology; Phase; Physicians; Pre-Clinical Model; Predisposition; Proto-Oncogene Protein c-kit; Publications; Regimen; Research; Research Methodology; Research Personnel; Research Technics; Residual state; Ribosomes; Risk; Role; Scientist; Second Primary Cancers; Sickle Cell; Signal Transduction; Structure; Testing; Therapeutic; Thrombopoietin; Time; Tissues; Toxic effect; Toxin; Training; Translational Research; Transplantation; Transplantation Conditioning; Universities; Whole-Body Irradiation; Work; Xenograft procedure; analytical method; antibody-dependent cell cytotoxicity; base; career; career development; cellular imaging; chemotherapy; chimeric antigen receptor T cells; conditioning; cytotoxicity; efficacy evaluation; experimental study; gene therapy; genotoxicity; in vivo; insight; irradiation; minimal risk; mortality; mouse model; novel; pre-clinical; receptor; skills; stem cell biology; stem cell engraftment; targeted agent; therapeutic development; transcriptome; transplantation therapy

Project Summary Hematopoietic stem cell transplant (HSCT) and gene therapy are increasingly utilized for definitive management of disorders affecting the lymphohematopoietic compartment. Conditioning regimens are one of the critical components HSCT. The majority of conditioning regimens used today are either based on alkylating chemotherapy, such as busulfan, or involve varying doses of total body irradiation (TBI). These agents are genotoxic and affect organs other than the hematopoietic compartment, leading to myriad acute and long-term HSCT associated complications. Our overarching goal is to develop a non-genotoxic conditioning regimen targeting hematopoietic stem and progenitor cells (HSPCs). Preliminary data from our lab indicates MPL (thrombopoietin receptor), a critical survival signal in hematopoietic stem cells (HSCs), could be an ideal candidate for development of HSPC-targeted HSCT conditioning approaches. Restricted expression profiles within the hematopoietic compartment, with a higher expression in HSC when compared to the hematopoietic progenitor compartment, make MPL a very attractive target. We have developed two strategies to target the MPL expressing HSPC compartment. First, an immunotoxin approach, generated by conjugating MPL antibody to a ribosomal toxin, saporin (MPL-ab-SAP), and the second, a MPL receptor engaging thrombopoietin based CAR-T cell (TPO-CAR-T cells). We hypothesize that a conditioning agent targeting the MPL receptor will result in selective HSPC depletion thereby creating adequate niche space to allow donor engraftment and will have minimal risk of cytotoxicity outside the hematopoietic compartment. We will determine the mechanism, and critical determinants of MPL-ab-SAP-mediated cytotoxicity in the HSC compartment. Then, we will determine whether HSPC depletion by MPL-ab-SAP could be utilized as an effective conditioning strategy for HSCT and autologous HSC-based gene therapy. The role of TPO-CAR-T cells as an HSCT conditioning will also be studied. The K08 Award will be used to protect my time for research, help broaden my research skills and knowledge, and provide an outstanding basis for a career in basic and translational HSCT and gene therapy. This mentored phase of training will allow me to further integrate and build on my research skills as a physician-scientist. Over the next five years, I will (1) gain expertise in molecular methods needed for development of therapeutic antibodies, and gene therapy based therapeutics, (2) gain further exposure in HSC biology, transplant immunology, and murine HSCT, (3) acquire skills in cell imaging and bioinformatics, (4) acquire skills in translational research methodology, grantsmanship and leadership, and (5) build on my publication record and develop research ideas for independent investigator grants. To accomplish these goals, I have developed a well-structured training plan, and I have assembled a highly-qualified mentorship team of Dr. Spencer (expertise in cell and gene therapy), Dr. Bunting (HSC biology), Dr. Waller (HSPC biology and clinical transplantation), and Dr. Larsen (transplant immunology) at Emory University.

项目摘要 造血干细胞移植（HSCT）和基因治疗越来越多地用于确定管理 影响淋巴瘤室的疾病。调理方案是关键的一种 组件HSCT。当今使用的大多数调节方案要么基于烷基化 化学疗法，例如Busulfan或涉及各种剂量的全身照射（TBI）。这些代理是 除造血室以外的遗传毒性和影响器官，导致无数急性和长期 HSCT相关并发症。我们的总体目标是制定一种非基因毒性调节方案 靶向造血干和祖细胞（HSPC）。来自我们实验室的初步数据指示MPL （血小板蛋白受体）是造血干细胞中的关键存活信号（HSC），可能是理想的 候选HSPC靶向HSCT调节方法的候选人。受限的表达曲线 在造血区室内，与造血相比，HSC的表达较高 祖细胞室，使MPL成为非常有吸引力的目标。我们已经制定了两种策略来针对 MPL表达HSPC室。首先，通过结合MPL抗体产生的免疫毒素方法 到核糖体毒素，糖果蛋白（MPL-AB-SAP），第二个MPL受体与基于血小板的基于血小板的受体 CAR-T细胞（TPO-CAR-T细胞）。我们假设针对MPL受体的调节剂将导致 在选择性HSPC耗竭中，从而创造了足够的利基空间以允许捐赠者植入，并将有 在造血室外的细胞毒性风险最小。我们将确定机制，并 HSC室中MPL-AB-SAP介导的细胞毒性的关键决定因素。然后，我们将确定 MPL-AB-SAP的HSPC耗竭是否可以用作HSCT和 基于HSC的自体基因疗法。还将研究TPO-CAR-T细胞作为HSCT调节的作用。 K08奖将用于保护我的研究时间，帮助扩大我的研究技能和知识， 并为基本和翻译HSCT和基因疗法的职业提供了出色的基础。这是指导的 培训阶段将使我能够进一步整合并以医生科学家的研究技巧为基础。超过 接下来的五年，我将（1）获得治疗开发所需的分子方法的专业知识 抗体和基于基因治疗的疗法，（2）在HSC生物学，移植中获得进一步的暴露 免疫学和鼠类HSCT，（3）获取细胞成像和生物信息学方面的技能，（4）获取技能 翻译研究方法论，授予技巧和领导力，以及（5）在我的出版记录和 为独立调查员赠款开发研究思想。为了实现这些目标，我已经开发了 结构良好的培训计划，我组建了Spencer博士的高度合格的指导团队（专业知识 在细胞和基因疗法中），邦廷博士（HSC生物学），沃勒博士（HSPC生物学和临床移植）和 埃默里大学（Emory University）Larsen博士（移植免疫学）。