Acute Hepatic Porphyrias: Pathogenesis & Treatment

急性肝卟啉症：发病机制

• 批准号: 8721940
• 负责人: Makiko Yasuda
• 金额: $ 15万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8721940
• 关键词: Acids; Acute; Acute Intermittent Porphyria; Adult; Affect; Aminolevulinic Acid; Autologous; Biochemical; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Brain; Carbon Tetrachloride; Cell Proliferation; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Defect; Development; Disease; Doctor of Philosophy; Dose; Effectiveness; Engraftment; Environment; Enzymes; Erythropoietic Porphyria; Evaluation; Family; Funding; Future; Gene Expression; Genes; Genetic; Genome; Genomics; Goals; Heme; Hepatic; Hepatic Porphyrias; Hepatocellular Damage; Hepatocyte; Human; Hydroxymethylbilane Synthase; Immunocompromised Host; Immunohistochemistry; Injection of therapeutic agent; Injury; Institutes; Intraperitoneal Injections; Journals; Knock-in Mouse; Laboratories; Laboratory Research; Lead; Lentivirus Vector; Life; Liver; Liver diseases; Luciferases; Mass Spectrum Analysis; Mentored Research Scientist Development Award; Mentors; Modeling; Molecular; Mus; Mutation; National Institute of Diabetes and Digestive and Kidney Diseases; Neurologic; Normal Cell; Pathogenesis; Pathway interactions; Patients; Performance; Phenobarbital; Phenotype; Physicians; Plasma; Porphobilinogen; Porphyrias; Porphyrins; Printing; Process; Property; Protocols documentation; Quality of life; Recurrence; Research; Research Personnel; Research Project Grants; Research Proposals; Residual state; Science; Scientist; Stem cells; Tail; Techniques; Technology; Therapeutic Studies; Training; Transgenic Mice; Translating; Translational Research; Transplantation; United States National Institutes of Health; Veins; Writing; base; bioluminescence imaging; career; career development; clinical phenotype; design; effective therapy; enzyme deficiency; erythroid differentiation; foot; gene therapy; heme 1; heme biosynthesis; hepatocyte engraftment; immunosuppressed; improved; in vivo; induced pluripotent stem cell; intrahepatic; knock-down; liquid chromatography mass spectrometry; liver cell proliferation; meetings; mouse model; non-oncogenic; novel; overexpression; prevent; professor; programs; public health relevance; research study; stem; stem cell biology; stem cell technology; transcriptome sequencing; urinary; vector

DESCRIPTION (provided by applicant): The overall goal of this NIDDK Mentored Research Scientist Development Award is to provide an integrated research and educational program for the applicant that will lead to an independent academic career in translational research. Robert J. Desnick, PhD, MD, Professor and Chair of Genetics and Genomic Sciences and Associate Dean for Genome- Based Research, and Ihor Lemischka, PhD, Professor of Gene and Cell Therapy and Director of the Black Family Stem Cell Institute, will serve as Mentor and Co-Mentor, respectively. This applicant will devote 95% of her effort to the career development program which includes: 1) undertaking the proposed laboratory research, 2) training in stem cell biology and iPS techniques, 3) participation in Genetics and Stem Cell Biology "Work in Progress" sessions, journal clubs and seminars, and attendance at national scientific meetings, and 4) bi-weekly meetings with Mentors. The specific aims of the proposed research are: 1) to investigate the pathogenesis of the life-threatening acute neurological attacks in Acute Intermittent Porphyria (AIP), the most common hepatic porphyria [due to deficient activity of hydroxymethylbilane synthase (HMB-synthase)], and 2) to evaluate the effectiveness of hepatocyte and bone marrow transplantation for AIP as proof-of-concept for future induced pluripotent stem (iPS) cell-based therapies for patients with AIP and the other acute hepatic porphyrias who have recurrent attacks. Towards this goal, Aim 1 will investigate disease pathogenesis in the previously generated T1/T2 AIP knock-down mice that have an "inducible" biochemical phenotype and in the newly generated AIP knock-in mice (R167Q+/+) that have a severe biochemical and clinical phenotype. Aim 2 will evaluate the effectiveness of primary hepatocyte transplantation and bone marrow transplantation in clearing the accumulated plasma and urinary porphyrin precursors, 4-aminolevulenic acid (ALA) and porphobilinogen (PBG) in the severely affected R167Q+/+ mice. These studies will determine the number of HMB-synthase competent cells necessary to clear the circulating porphyrin precursors and whether the clearance also down-regulates hepatic 5'- aminolevulinic acid synthase 1 (ALAS1) activity. Aim 3 will assess whether transplanting HMB-synthase overexpressing hepatocytes and bone marrow (transduced ex vivo with lentiviral vectors prior to transplantation) can effectively decrease the number of engrafted cells required to clear ALA and PBG accumulation in the R167Q+/+ mice. This is of particular importance, as it is a major challenge to achieve high levels of donor cell engraftment and proliferation for diseases such as AIP that lack hepatocellular damage and thus do not provide the donor cells a proliferative advantage. Based on the results of Aims 2 and 3, Aim 4 will apply the latest iPS technology to develop treatment for the AIP mice as a model for future applications in patients. The rationale for cell transplantation to prevent the acute attacks in AIP is supported by recent studies in our laboratory demonstrating that intraperitoneal injection of an AAV2/8 vector overexpressing HMB-synthase effectively and continuously prevented the biochemical induction of an acute attack in the T1/T2 mouse model of AIP (see Preliminary Results, Section C.3.). This integrated research and educational program, together with the outstanding research environment at Mount Sinai, should facilitate the applicant's transition to an academic career as an independent translational researcher.

描述（由申请人提供）：NIDDK 指导研究科学家发展奖的总体目标是为申请人提供综合的研究和教育计划，从而在转化研究领域实现独立的学术生涯。遗传学和基因组科学教授兼系主任、基因组研究副院长 Robert J. Desnick 博士、医学博士和基因与细胞治疗教授兼 Black Family 干细胞研究所所长 Ihor Lemischka 博士将分别担任导师和联合导师。该申请人将把 95% 的精力投入到职业发展计划中，其中包括：1) 进行拟议的实验室研究，2) 干细胞生物学和 iPS 技术培训，3) 参加遗传学和干细胞生物学“正在进行的工作”会议、期刊俱乐部和研讨会，以及参加国家科学会议，以及 4) 每两周一次与导师的会议。本研究的具体目的是：1) 调查急性间歇性卟啉症 (AIP) 中危及生命的急性神经系统发作的发病机制，这是最常见的肝卟啉症 [由于羟甲基胆烷合酶 (HMB-合酶) 活性缺陷]，2) 评估肝细胞和骨髓移植的有效性 AIP 作为未来基于诱导多能干 (iPS) 细胞的疗法的概念验证，用于治疗 AIP 和其他反复发作的急性肝卟啉症患者。为了实现这一目标，Aim 1 将研究先前生成的具有“可诱导”生化表型的 T1/T2 AIP 敲除小鼠和新生成的具有严重生化和临床表型的 AIP 敲入小鼠 (R167Q+/+) 的疾病发病机制。目标 2 将评估原代肝细胞移植和骨髓移植在清除严重受影响的 R167Q+/+ 小鼠中累积的血浆和尿卟啉前体、4-氨基乙酰丙酸 (ALA) 和胆色素原 (PBG) 方面的有效性。这些研究将确定清除循环卟啉前体所需的 HMB 合酶感受态细胞的数量，以及清除是否也会下调肝脏 5'-氨基乙酰丙酸合酶 1 (ALAS1) 活性。目标 3 将评估移植过表达 HMB 合酶的肝细胞和骨髓（移植前用慢病毒载体离体转导）是否可以有效减少清除 R167Q+/+ 小鼠中 ALA 和 PBG 积累所需的移植细胞数量。这是特别重要的，因为对于诸如 AIP 等缺乏肝细胞损伤的疾病，实现高水平的供体细胞植入和增殖是一个重大挑战，这些疾病缺乏肝细胞损伤，因此不能为供体细胞提供增殖优势。基于Aims 2和3的结果，Aim 4将应用最新的iPS技术来开发AIP小鼠的治疗方法，作为未来在患者中应用的模型。我们实验室最近的研究支持了细胞移植预防 AIP 急性发作的基本原理，该研究表明，腹膜内注射过表达 HMB 合酶的 AAV2/8 载体可有效且持续地阻止 AIP T1/T2 小鼠模型中急性发作的生化诱导（参见初步结果，C.3 节）。这项综合研究和教育计划，加上西奈山出色的研究环境，应有助于申请人过渡到作为独立转化研究员的学术生涯。