Hepatocyte Transplantation for Liver Based Metabolic Disease

肝细胞移植治疗肝脏代谢疾病

• 批准号: 9762098
• 负责人: Ira J. Fox
• 金额: $ 95.84万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-10 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9762098
• 关键词: Acute; Adolescent; Adult; Allografting; Amino Acids; Ammonia; Animal Model; Animals; Autologous; Bilirubin; Biological Assay; Biology; Blood; Carbon Dioxide; Cell Transplantation; Cell Transplants; Cells; Child; Clinic; Clinical; Development; Diagnosis; Diet; Disease; Disease model; Engraftment; Enrollment; Event; FDA approved; Fibroblasts; Gene Delivery; Generations; Goals; Graft Rejection; Hepatic; Hepatocyte; Hepatocyte transplantation; Immune; Immunologic Monitoring; Immunology procedure; Immunosuppression; Incidence; Intestines; Isomerism; Isotopes; Kidney; Laboratories; Liver; Liver diseases; Measures; Metabolic; Metabolic Diseases; Metabolism; Modeling; Monitor; Motivation; Neonatal Screening; Neurologic; Neurologic Dysfunctions; Neurologic Symptoms; Oral; Organ Transplantation; Palate; Pathologic Processes; Pathway interactions; Patients; Peripheral Blood Lymphocyte; Phenylalanine Hydroxylase; Phenylketonurias; Pilot Projects; Process; Radiation; Reporting; Risk; Safety; Solid; Stem cells; Stimulus; System; T-Lymphocyte; TNFSF5 gene; Techniques; Technology; Testing; Therapeutic; Time; Transplant Recipients; Transplantation; base; carbon skeleton; conditioning; executive function; experience; experimental study; fatty acid oxidation; gene transplantation for gene therapy; graft function; immunoreactivity; in vivo; indexing; induced pluripotent stem cell; innovation; liver transplantation; minimally invasive; neuropsychiatry; non-invasive monitor; nonhuman primate; novel; organic acid; oxidation; parenteral administration; patient population; population based; standard measure; success; targeted treatment; vector; young adult

Transplantation of isolated parenchymal cells has great promise as a minimally invasive therapy and partial success has been reported for several rare metabolic liver diseases. Major limitations to the technique include an inability to transplant an adequate mass of donor hepatocytes, and lack of a sensitive enough technology to monitor the status of transplanted cells. This proposal includes several major innovations in understanding hepatocyte transplant biology and optimizing its application. In animal models liver-directed radiation facilitates repopulation of the native liver by transplanted hepatocytes. Unfortunately, the standard clinical and laboratory signs that identify rejection in solid organ transplantation are not useful in recipients of cell transplants, and the sensitivity of functional changes following hepatocyte transplant is inadequate to diagnose rejection before damage to the allograft is irreversible. The availability of assays that would allow non-invasive monitoring of graft function and assessment of rejection risk would facilitate more rational management of patients following hepatocyte transplantation. As diseases targeted for treatment by hepatocyte transplantation are rare, and most reports involve anecdotal experience in a diverse patient population, we will transplant patients with phenylketonuria (PKU), a disease with an incidence that will make adequate patient enrollment possible. Phenylalanine hydroxylase (PAH) deficiency, traditionally known as PKU was the original motivation for population-based newborn screening. However, standard dietary management strategies have failed in older adolescents and adults due to difficulty in adhering to diet, leading to diminished executive function, and other neurologic and neuropsychiatric problems. The proposal has three specific aims. Aim 1 is to confirm the safety of liver-directed radiation conditioning and document 5-10% replacement of host hepatocytes by donor hepatocytes. We hypothesize that use of multiple donors and pre-transplant hepatic radiation conditioning will lead to competitive repopulation of the host liver and clinical correction of PAH deficiency in patients with PKU, a model disease for treatment of liver-based metabolic diseases. Aim 2 is to identify graft rejection by immunologic monitoring and to successfully treat it before the process is irreversible. We hypothesize that monitoring rejection risk by a proven assay of donor-specific T cell immune reactivity can be used to assess the adequacy of immune suppression following cell transplantation. Aim 3 is to determine the extent to which use of real-time measures of donor hepatocyte function, rather than use of surrogates, will allow assessment of graft function after hepatocyte transplantation. We hypothesize that isotopic Phe turnover studies in PKU patients can more effectively measure changes in Phe metabolism than standard measures. These results will be important for the possible transplantation of gene edited autologous or stem cell-derived hepatocytes. Delivery of gene editing vectors ex vivo to hepatocytes is significantly more efficient than in vivo delivery, and is even more efficient in patient-derived fibroblasts that would later be converted to iPS-derived hepatocytes.

分离的实质细胞移植作为一种微创疗法和部分治疗具有广阔的前景。 据报道，该疗法在治疗几种罕见的代谢性肝病方面取得了成功。该技术的主要限制包括 无法移植足够量的供体肝细胞，并且缺乏足够敏感的技术来移植 监测移植细胞的状态。该提案包括在理解上的几项重大创新 肝细胞移植生物学及其应用优化。在动物模型中，肝脏定向辐射有利于 通过移植的肝细胞重建原生肝脏。不幸的是，标准的临床和实验室 实体器官移植中识别排斥反应的迹象对于细胞移植受者来说是没有用的，并且 肝细胞移植后功能变化的敏感性不足以诊断排斥反应 同种异体移植物的损伤是不可逆的。允许非侵入性监测的检测方法的可用性 移植物功能和排斥风险评估将有助于对术后患者进行更合理的管理 肝细胞移植。由于肝细胞移植治疗的靶向疾病很少见，并且 大多数报告涉及不同患者群体的轶事经历，我们将移植患者 苯丙酮尿症（PKU），这种疾病的发病率足以使足够的患者入组成为可能。 苯丙氨酸羟化酶 (PAH) 缺乏症（传统上称为 PKU）是 基于人群的新生儿筛查。然而，标准的饮食管理策略在老年人中却失败了。 青少年和成人由于难以坚持饮食，导致执行功能下降，以及其他 神经系统和神经精神问题。该提案有三个具体目标。目标1是确认安全性 肝脏定向辐射调理并记录供体肝细胞 5-10% 的替换 肝细胞。我们假设使用多个供体和移植前肝脏辐射调节将 导致宿主肝脏的竞争性再生和 PKU 患者 PAH 缺陷的临床纠正， 治疗肝脏代谢疾病的模型疾病。目标 2 是通过以下方式识别移植物排斥反应： 免疫监测并在该过程不可逆转之前成功治疗它。我们假设 通过经过验证的供体特异性 T 细胞免疫反应测定来监测排斥风险，可用于评估 细胞移植后免疫抑制的充分性。目标 3 是确定 使用供体肝细胞功能的实时测量，而不是使用替代物，将允许评估 肝细胞移植后的移植物功能。我们假设 PKU 中的同位素 Phe 周转研究 与标准测量相比，患者可以更有效地测量 Phe 代谢的变化。这些结果将 对于基因编辑的自体或干细胞衍生的肝细胞的可能移植很重要。 基因编辑载体离体递送至肝细胞的效率明显高于体内递送，并且 在患者来源的成纤维细胞中甚至更有效，这些成纤维细胞随后会转化为 iPS 来源的肝细胞。