Improving Islet Transplantation Outcome With Akt1

使用 Akt1 改善胰岛移植结果

基本信息

批准号：
8049189
负责人：
HONGJU WU
金额：
$ 31.23万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8049189
关键词：
1-Phosphatidylinositol 3-Kinase Adenoviruses Adverse effects Apoptosis Autoimmune Process Candidate Disease Gene Capsid Cell Proliferation Cell Survival Cells Clinic Clinical Research Dose Employment Exhibits Gene Delivery Gene Expression Gene Transfer Genes Growth Factor Health Herpesvirus 1 Human Immune response Induction of Apoptosis Infection Inflammation Insulin Insulin-Dependent Diabetes Mellitus Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Knockout Mice Malignant - descriptor Malignant Neoplasms Mediating Methods Modality Modification Names Outcome Pancreas Pathway interactions Patients Process Proliferating Protein-Serine-Threonine Kinases Proto-Oncogene Proteins c-akt Public Health Rattus Research Personnel Rodent Safety Serotyping Signal Transduction Pathway TK Gene Therapeutic Thymidine Kinase Time Toxic effect Transgenes Transgenic Mice Translating Transplantation Viral Viral Vector base clinically significant imaging modality improved in vivo interest islet mouse model promoter success suicide gene transplantation typing treatment strategy vector

1-Phosphatidylinositol 3-Kinase Adenoviruses Adverse effects Apoptosis Autoimmune Process Candidate Disease Gene Capsid Cell Proliferation Cell Survival Cells Clinic Clinical Research Dose Employment Exhibits Gene Delivery Gene Expression Gene Transfer Genes Growth Factor Health Herpesvirus 1 Human Immune response Induction of Apoptosis Infection Inflammation Insulin Insulin-Dependent Diabetes Mellitus Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Knockout Mice Malignant - descriptor Malignant Neoplasms Mediating Methods Modality Modification Names Outcome Pancreas Pathway interactions Patients Process Proliferating Protein-Serine-Threonine Kinases Proto-Oncogene Proteins c-akt Public Health Rattus Research Personnel Rodent Safety Serotyping Signal Transduction Pathway TK Gene Therapeutic Thymidine Kinase Time Toxic effect Transgenes Transgenic Mice Translating Transplantation Viral Viral Vector base clinically significant imaging modality improved in vivo interest islet mouse model promoter success suicide gene transplantation typing treatment strategy vector

展开

项目摘要

DESCRIPTION (provided by applicant): Summary Islet transplantation is becoming a potential cure for type 1 diabetes (T1D). However, the limited supply and significant islet loss in the peritransplant period are cast as the major limitations of this treatment strategy. This project is aimed to improve the therapeutic outcome of islet transplantation by introducing constitutively active Akt1 (CA-Akt1) into the insulin producing 2-cells ex vivo. The serine/threonine protein kinase Akt/PKB is the direct downstream target of PI3 Kinase pathway, and has been found to have dual functions of anti-apoptosis and induction of cell proliferation. Relevance of Akt on 2-cell survival and proliferation has been demonstrated in studies of transgenic and knockout mouse models, as well as using pharmacological methods. Nonetheless, in order to realize the therapeutic potential of CA-Akt1, a safe, efficient and specific vector is needed to deliver Akt1 into islet 2-cells ex vivo. In this regard, adenovirus serotype 5 (Ad5)-based vector is of great interest. Our previous studies have demonstrated the modified Ad5 vector, AdRGDpK7, exhibited significantly higher gene transfer efficiency for the human islet cells. We thus propose to employ Ad5RGDpK7 to deliver Akt1 into islet cells ex vivo. To further diminish the potential adverse effect of CA-Akt1, we will restrict exogenous Akt1 expression in 2-cells by employment of 2-cell specific promoter-rat insulin promoter (RIP) to drive Akt1 expression. In addition, we propose to co-express a dual functional modality, HSV-TK, with CA-Akt1 so that it can be used as both a non-invasive imaging modality to follow the transplanted islets and a suicide gene should malignancy occur. Our specific aims are thus: 1) To develop a 2-cell specific, infectivity-enhanced Ad5 vector that allows efficient and specific CA-Akt1 and HSV-TK gene delivery into 2-cells ex vivo; 2) To examine the capacity of the Ad5 vector developed above to promote islet survival and proliferation while minimizing transformation, thus enhancing the efficacy of islet transplantation; and 3) To evaluate the safety of the Ad5 vector developed above in the context of islet transplantation. PUBLIC HEALTH RELEVANCE: It is clear that islet transplantation holds great potential for the cure of Type 1 Diabetes. This study seeks to improve the therapeutic outcome of islet transplantation by modifying the islets with protective genes using a highly efficient, specific and safe gene delivery vector. Success of this study is expected to have significant impact in the field of islet transplantation treatment for type 1 diabetes.

描述（由申请人提供）：摘要胰岛移植已成为1型糖尿病（T1D）的潜在治愈方法。但是，在本植物期间，有限的供应和大量胰岛损失被认为是该治疗策略的主要局限性。该项目的目的是通过将组成性活跃的AKT1（CA-AKT1）引入产生2细胞的胰岛素后，通过将胰岛移植的治疗结果提高。丝氨酸/苏氨酸蛋白激酶AKT/PKB是PI3激酶途径的直接下游靶标，已发现具有抗凋亡的双重功能和细胞增殖的诱导。在转基因和基因敲除小鼠模型的研究以及使用药理学方法中，AKT对2细胞存活和增殖的相关性已证明。尽管如此，为了实现CA-AKT1的治疗潜力，需要安全，高效和特定的向量将AKT1传递到离体2细胞中。在这方面，基于腺病毒的血清型5（AD5）载体引起了极大的兴趣。我们以前的研究表明，经过修改的AD5载体ADRGDPK7在人类胰岛细胞中表现出明显更高的基因转移效率。因此，我们建议使用AD5RGDPK7将Akt1输送到离体的胰岛细胞中。为了进一步减少CA-AKT1的潜在不利影响，我们将通过使用2细胞特异性启动子rat胰岛素启动子（RIP）来驱动AKT1表达，从而限制2细胞中的外源AKT1表达。此外，我们建议与CA-AKT1共同表达双重功能模态HSV-TK，以便可以用作非侵入性成像方式，以遵循移植的胰岛和自杀基因，应发生恶性肿瘤。因此，我们的具体目的是：1）开发一个2细胞特异性，感染性增强的AD5载体，该载体可以使有效的，特定的Ca-Akt1和HSV-TK基因递送到2个细胞中； 2）检查上面开发的AD5载体的能力以促进胰岛存活和增殖，同时最大程度地减少转化，从而增强了胰岛移植的功效； 3）评估在胰岛移植的背景下在上面开发的AD5矢量的安全性。公共卫生相关性：很明显，胰岛移植具有巨大的治疗1型糖尿病的潜力。这项研究旨在通过使用高效，特定和安全的基因递送载体修饰胰岛通过保护性基因来改善胰岛移植的治疗结果。预计这项研究的成功将在1型糖尿病的胰岛移植治疗领域产生重大影响。