Engineering pancreatic islets with TGβ protein to overcome rejection

用 TGβ 蛋白改造胰岛以克服排斥反应

基本信息

批准号：
8966928
负责人：
Haval Shirwan
金额：
$ 22.5万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-12 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8966928
关键词：
Affect Affinity Alloantigen Allogenic Antigens Applications Grants Area Autoantigens Autoimmune Diseases Autoimmune Process Autoimmunity Beta Cell Biological Biotin C57BL/6 Mouse Cells Characteristics Chimeric Proteins Chronic Clinical Data Development Diabetes Mellitus Diabetic mouse Disease Economic Burden Ectopic Expression Engineering Failure Family suidae Fibrosis Generations Goals Graft Rejection Graft Survival Health Homeostasis Human IL2RA gene Immune Immune response Immune system Immunosuppression Immunosuppressive Agents Inbred BALB C Mice Inbred NOD Mice Insulin Insulin-Dependent Diabetes Mellitus Intervention Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Kidney Ligands Maintenance Mediating Membrane Memory Methods Modeling Molecular Molecular Chaperones Morbidity - disease rate Mus Natural regeneration Pancreas Pancreas Transplantation Pharmaceutical Preparations Play Population Proteins Protocols documentation Recovery Regulation Regulatory T-Lymphocyte Reporting Rodent Model Role Safety Series Signal Transduction Sirolimus Source Splenocyte Streptavidin Suggestion Surface System T-Lymphocyte Therapeutic To autoantigen Toxic effect Transforming Growth Factor beta Transgenic Organisms Transplantation Viral Vector base clinically relevant crosslink design diabetic effective therapy efficacy testing gene therapy human disease immune function immunoregulation improved functioning islet lymph nodes macrophage mortality mouse model novel novel strategies pleiotropism prevent receptor standard care

项目摘要

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) continues to be a major source of morbidity and mortality in over 0.9% US population with an economic burden in billions of dollars. The two current methods used to treat T1D are insulin therapy and allogeneic islet/pancreas transplantation (Tx), both of which have major limitations. Insulin administration does not completely prevent the complications associated with diabetes. Allogeneic islet/pancreas Tx suffers from rejection as well as renal and islet toxicities associated with chronic use of immunosuppressive drugs. Therefore, the primary objective of this grant proposal is to establish a novel immunomodulatory protocol to confer tolerance to allogeneic islet grafts in the absence of continuous immunosuppression. This will be achieved by engineering allogeneic islets ex vivo to transiently display on their surface a novel form of TGFß1, SA-TGFß, protein and use the engineered islets to induce tolerance and treat T1D using rodent models. T1D and allogeneic islet cell destruction is primarily mediated by T cells directed at unique beta cell and Tx antigens. Controlling the function of T pathogenic cells is, therefore, critical to tolerance induction to auto/alloantigens and treatment of T1D using allogeneic islets. We herein propose to use SA-TGFß as an immunomodulatory molecule to achieve this goal. TGFß1 has pleiotropic immune functions and is critical for tolerance to self-antigen. TGFß also plays an important role in acquired tolerance to auto and alloantigens using various immunomodulatory approaches. Most importantly, ectopic expression of TGFß in pancreatic islets using transgenic systems or viral vectors proved effective in inducing tolerance to autoantigens in NOD mouse model of human T1D. However, continuous expression of this molecule is associated with massive fibrosis and long-term beta cell failure. Importantly, transient expression of TGFß in beta cells using inducible transgenic systems overcame its deleterious effects without compromising its tolerogenic function. Inasmuch as this transgenic approach is not applicable to clinical islet Tx, we envisioned that the transient display of TGFβ at protein level on the surface of islets in a clinically applicable manner may achieve tolerance without its reported complications arising from continuous expression. In limited preliminary studies, we demonstrated that the transient display of SA-TGFß on pancreatic islets was effective in overcoming rejection in a chemically induced islet Tx model. Building on these strong preliminary data, we herein propose to use SA-TGFß-engineered allogeneic islets as a means of immunomodulation to prevent graft rejection and treat diabetes in chemically and spontaneously diabetic mouse models. A series of studies will be conducted to delineate the implicated mechanisms of the tolerance. Rapid and transient display of SA-TGFß protein on pancreatic islets offers a whole new means of intervention in the areas of autoimmunity and Tx. This approach possesses the simplicity, safety, and efficacy required to make it a clinically relevant alternative that may accomplish the same goals as gene therapy in the treatment of a broad spectrum of immune-based diseases, including T1D.

描述（由适用提供）：1型糖尿病（T1D）仍然是美国超过0.9％人口的发病率和死亡率的主要来源，经济燃烧数十亿美元。用于治疗T1D的两种当前方法是胰岛素治疗和同种异体胰岛/胰腺移植（TX），这两种方法都有主要的局限性。胰岛素给药并不能完全防止与糖尿病有关的并发症。同种异体胰岛/胰腺TX遭受与长期使用免疫抑制药物有关的排斥和肾脏和胰岛毒性。因此，该赠款提案的主要目的是建立一种新颖没有连续的免疫抑制。这将通过工程同种异体胰岛实行，以瞬时在其表面上显示出一种新型TGFß1，SA-TGFß，蛋白质的新形式，并使用工程的胰岛使用啮齿动物模型来诱导耐受性和治疗T1D。 T1D和同种异体胰岛细胞破坏主要是由针对独特β细胞和TX抗原的T细胞介导的。因此，控制T致病细胞的功能对于使用同种异体胰岛对耐受性诱导自动/同种抗原和T1D处理至关重要。我们在这里提出，将SA-TGFß用作免疫调节分子来实现这一目标。 TGFß1具有多效免疫功能，对于耐受性抗原至关重要。 TGFß使用各种免疫调节方法在获得对汽车和同种抗原的耐受性中也起着重要作用。最重要的是，使用转基因系统或病毒载体在人类T1D的NOD小鼠模型中，使用转基因系统或病毒载体在胰岛中TGFß的生态表达被证明有效。但是，该分子的连续表达与大规模纤维化和长期β细胞衰竭有关。重要的是，使用诱导的转基因系统在β细胞中TGFß的瞬时表达克服了其有害作用，而不会损害其耐受性功能。由于这种转基因方法不适用于临床胰岛TX，因此我们设想，以临床适用的方式，TGFβ在胰岛表面上的TGFβ在蛋白质水平上的瞬时显示可能会达到公差，而不会导致其持续表达引起的并发症。在有限的初步研究中，我们证明了Sa-TGFß在胰岛上的瞬时显示可有效克服化学诱导的胰岛TX模型中的排斥反应。在这些强大的初步数据的基础上，我们在这里建议使用SA-TGFß工程化的同种异体胰岛作为免疫调节的一种手段，以防止成绩排斥和治疗化学和赞助的糖尿病和糖尿病小鼠模型。将进行一系列研究来描述公差的隐含机制。胰岛上SA-TGFß蛋白的快速和瞬时显示为自身免疫和TX领域的干预提供了全新的含义。这种方法具有使其成为临床上相关的替代方案所需的简单，安全性和效率，该替代方案可能在治疗包括T1D在内的广泛的免疫疾病中实现与基因治疗相同的目标。