The role of oxidative stress in the pathogenesis of Reticular Dysgenesis and thetherapeutic potential of antioxidants

氧化应激在网状发育不全发病机制中的作用和抗氧化剂的治疗潜力

• 批准号: 9317208
• 负责人: Katja Gabriele Weinacht
• 金额: $ 4.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-14 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9317208
• 关键词: Adenine Nucleotides; Adenosine Diphosphate; Adenosine Monophosphate; Advisory Committees; Affect; Aftercare; Antioxidants; Area; Award; Bacterial Infections; Biology; Boston; CRISPR/Cas technology; Cause of Death; Cell Differentiation process; Cell Line; Cell model; Cell physiology; Cessation of life; Child; Clinical; Complement; Constitution; Dana-Farber Cancer Institute; Data; Defect; Development; Development Plans; Diagnosis; Disease; Disease model; Effectiveness; Engraftment; Ensure; Enzymes; Generations; Genes; Glutathione; Goals; Health; Hematological Disease; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Immune; Immunologic Deficiency Syndromes; Immunology; In Situ; In Vitro; Institution; Lead; Life; Lymphopenia; Mediating; Mentors; Metabolic; Metabolism; Microbiology; Mitochondria; Modeling; Molecular; Mus; Mutate; Mutation; Mycoses; Myelogenous; Myeloid Cells; Natural Immunity; Nature; Neutropenia; Oncology; Outcome; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Pathology; Patients; Pediatric Hematologist; Pediatric Hematology; Pediatric Hospitals; Physicians; Play; Positioning Attribute; Research; Research Personnel; Research Project Grants; Research Proposals; Research Training; Role; Scientist; Sensorineural Hearing Loss; Severe Combined Immunodeficiency; Specimen; Staging; Structure; Supportive care; Testing; Therapeutic; Therapeutic Human Experimentation; Time; Training; Translating; Translational Research; Transplantation; Universities; Work; Xenograft Model; Xenograft procedure; Zebrafish; adaptive immunity; adenylate kinase; base; career; career development; clinical care; congenital immunodeficiency; design; experience; gene correction; high risk; human disease; humanized mouse; improved; in vitro Model; in vivo; induced pluripotent stem cell; infancy; mitochondrial dysfunction; mitochondrial metabolism; mouse model; neutrophil; novel strategies; outcome forecast; progenitor; reconstitution; reticular dysgenesis; stem; stem cell biology; stem cell differentiation; stem cells; tandem mass spectrometry; targeted treatment; tenure track; therapeutic target; tissue culture

 DESCRIPTION (provided by applicant): Reticular Dysgenesis (RD) is one of the most serious forms of severe combined immunodeficiency (SCID) because it affects both innate and adaptive immunity. The disease is characterized by arrested neutrophil maturation, profound lymphopenia, and sensorineural hearing loss. It is invariably fatal early in life unless immune reconstitution is achieved by hematopoietic stem cell transplantation (HSCT). The simultaneous occurrence of severe neutropenia and lymphopenia is responsible for a high risk of death in infancy, and the predominance of bacterial and fungal infections supports neutropenia as the prevailing cause of death. RD is caused by mutations in the mitochondrial enzyme Adenylate Kinase 2 (AK2), however, how AK2 defects translate into disease pathology is largely unknown. Overall, transplant outcomes in RD are significantly worse compared to any other form of SCID, suggesting that the nature of the genetic defect may directly impact the poor prognosis. Therefore, elucidating the mechanistic basis of RD is critical in order to target the underlying problem and develop additional therapeutic options. Our prior work in induced pluripotent stem cell and zebrafish models of RD has shown that hematopoietic stem and progenitor cell differentiation is compromised but can be rescued by treatment with antioxidant agents (Rissone, Weinacht et al., J Exp Med, 2015, in press). These data led to the hypothesis that AK2 defects impair mitochondrial metabolism and increase oxidative stress, and that antioxidant agents improve mitochondrial function and represent a targeted supportive therapy to treat the neutropenia and overall constitution of patients with RD. To test this hypothesis I will dissect th molecular mechanisms underlying RD and their responsiveness to antioxidants by defining mitochondrial metabolism, oxidative stress and antioxidant reserve in different iPSC-models of RD before and after treatment with antioxidants. To demonstrate that the in vitro findings can be translated to patients, I will develop a xenograft model of RD based on transplantation of AK2-deficient iPSC-derived respecified multipotent hematopoietic progenitors into NSG-mice, and examine how antioxidant treatment affects engraftment, differentiation potential and mitochondrial function. If my preliminary findings that antioxidants improve mitochondrial function are supported by the proposed research, the therapeutic potential of these agents could be exploited in a much wider range of diseases, in which mitochondrial pathology and oxidative stress are at play. I am a pediatric hematologist with focus on immune and immune-mediated diseases and substantial prior research experience in molecular microbiology, stem cell differentiation and reprogramming, who is seeking K08 support for mentored research under the guidance of Dr. Luigi Notarangelo, Division of Immunology, Boston Children's Hospital, with Dr. George Daley, Division of Hematology/Oncology, Boston Children's Hospital/Dana-Farber Cancer Institute, as co-mentor. My long-term career objective is to obtain a tenure-track position as a physician-scientist at an academic center with special commitment to the study, diagnosis and treatment of primary immunodeficiencies. The K08 award will provide the protected time I need to advance my training in stem cell biology, metabolism, xenografting, in situ gene editing and translational research. I will devote a minimum of 80% of my time to a focused research project investigating the molecular mechanisms underlying RD and its responsiveness to antioxidants, and will complement this with 20% of my effort dedicated to clinical care of children with hematologic diseases. Boston Children's Hospital, Dana-Farber Cancer Institute and Harvard University are internationally recognized research institutions with renowned expert researchers in the areas of stem cell biology, hematopoiesis, metabolism, in situ gene editing, and development of xenograft-models. Furthermore, the Divisions of Pediatric Hematology/Oncology and Immunology have a distinguished record of training successful physician-scientists. I have assembled a mentoring and advisory committee, consisting of Drs. Raif Geha, David Williams, Alan Beggs, and Suneet Agarwal, who will guide my research and training experience. The expertise of my advisory committee will be complemented by a set of additional collaborators who are experts in their respective fields (Dr. Kiran Musunuru, gene editing with CRISPR/Cas9; Dr. Marcia Haigis, mitochondrial biology; and Dr. Giancarlo la Marca, Tandem Mass Spectrometry). This research proposal is part of a structured plan with scientific, technical, clinical training and career development components. The career development plan builds upon my prior research and clinical experiences with the goal of ensuring that I acquire the expertise required to become a successful, independent investigator whose focus is on translating disease mechanisms underlying immunodeficiencies into targeted therapies for patients.

 描述(申请人提供)：网状发育不良(RD)是严重联合免疫缺陷(SCID)最严重的形式之一，因为它影响先天免疫和获得性免疫。这种疾病的特征是中性粒细胞成熟受阻，严重的淋巴细胞减少，以及感觉神经性听力损失。除非通过造血干细胞移植(HSCT)实现免疫重建，否则它在生命早期总是致命的。严重的中性粒细胞减少和淋巴细胞减少同时发生是婴儿期高死亡风险的原因，细菌和真菌感染的优势支持中性粒细胞减少症作为主要死亡原因。RD是由线粒体酶AK2突变引起的，然而，AK2缺陷如何转化为疾病病理在很大程度上尚不清楚。总体而言，与任何其他形式的SCID相比，RD患者的移植结果明显更差，这表明遗传缺陷的性质可能直接影响预后不良。因此，阐明RD的机制基础对于针对潜在问题和开发其他治疗方案至关重要。我们之前在诱导多能干细胞和斑马鱼RD模型中的工作表明，造血干细胞和祖细胞的分化受到损害，但可以通过抗氧化剂的治疗来挽救(Rissone，Weinacht等人，J Exp Med，2015年出版)。这些数据导致假设，AK2缺陷损害线粒体新陈代谢并增加氧化应激，抗氧化剂改善线粒体功能，并代表一种有针对性的支持性治疗，以治疗中性粒细胞减少症和RD患者的整体体质。为了验证这一假设，我将通过定义抗氧化剂治疗前后不同的RD IPSC模型中的线粒体代谢、氧化应激和抗氧化储备来剖析RD及其对抗氧化剂的反应性的分子机制。为了证明体外研究结果可以移植到患者身上，我将建立一个基于AK2缺乏的IPSC衍生的再特异性多潜能造血祖细胞移植到NSG-小鼠体内的RD异种移植模型，并检测抗氧化治疗如何影响植入、分化潜力和线粒体功能。如果我的初步发现--抗氧化剂改善线粒体功能--得到了拟议中的研究的支持，那么这些抗氧化剂的治疗潜力可以在更广泛的疾病中得到开发，在这些疾病中，线粒体病理和氧化应激起着作用。我是一名儿科血液学家，专注于免疫和免疫介导性疾病，在分子微生物学、干细胞分化和重编程方面有丰富的先前研究经验，正在寻求K08支持，在波士顿儿童医院免疫科Luigi Notarangelo博士的指导下，波士顿儿童医院/Dana-Farber癌症研究所血液/肿瘤科George Daley博士作为共同导师。我的长期职业目标是在一个学术中心获得一个终身医生兼科学家的职位，专门致力于初级免疫缺陷的研究、诊断和治疗。K08奖将为我在干细胞生物学、新陈代谢、异种移植、原位基因编辑和翻译研究方面的培训提供所需的保护时间。我会将至少80%的时间投入到一个有重点的研究项目中，研究RD背后的分子机制及其对抗氧化剂的反应，并将20%的精力用于儿童的临床护理。 患有血液病。波士顿儿童医院、Dana-Farber癌症研究所和哈佛大学是国际公认的研究机构，在干细胞生物学、造血、新陈代谢、原位基因编辑和异种移植模型开发等领域拥有著名的专家研究人员。此外，儿科血液科/肿瘤科和免疫科在培养成功的内科科学家方面有着杰出的记录。我已经组建了一个由Raif Geha博士、David Williams博士、艾伦·贝格斯博士和Suneet Agarwal博士组成的指导和咨询委员会，他们将指导我的研究和培训经验。我的咨询委员会的专业知识将得到其他一批各自领域的专家(CRISPR/Cas9基因编辑的Kiran Musunuru博士；线粒体生物学的Marcia Haigis博士；以及串联质谱仪的Giancarlo la Marca博士)的其他合作者的补充。这项研究提案是一项结构化计划的一部分，其中包括科学、技术、临床培训和职业发展部分。职业发展计划建立在我之前的研究和临床经验的基础上，目标是确保我获得所需的专业知识，成为一名成功的独立研究员，专注于将免疫缺陷潜在的疾病机制转化为患者的靶向治疗。