APOPTOSIS:A MEANS OF IMMUNE REGULATION TO TREAT DIABETES

细胞凋亡：治疗糖尿病的免疫调节手段

• 批准号: 6612650
• 负责人: Haval Shirwan
• 金额: $ 21.6万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-15 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6612650
• 关键词: B lymphocyte; T lymphocyte; apoptosis; autoantigens; autoimmunity; cysteine endopeptidases; cytokine receptors; diabetes mellitus therapy; gene expression; immunoregulation; insulin dependent diabetes mellitus; interleukin 2; laboratory mouse; pancreatic islets; tissue mosaicism

DESCRIPTION (provided by applicant): The main objective of this grant is to induce apoptosis specifically in auto and alloreactive lymphocytes by activating endogenous caspases. Physical elimination of these lymphocytes in NOD mouse is expected to abrogate the autoimmune process in prediabetic animals and to prevent islet allograft rejection in diseased animals, leading to the prevention and treatment of diabetes. Type I insulin-dependent diabetes mellitus (IDDM) is a chronic autoimmune disorder that affects a large number of people worldwide. Advances in understanding of the molecular and cellular basis of IDDM pathogenesis and improvements in medical practice have resulted in effective therapeutic approaches for the prevention of IDDM. Despite advances in medical therapies, IDDM remains a major cause of long-term morbidity and mortality. It is the single most common cause of renal failure requiring transplantation. Transplantation of whole pancreas or purified islets containing insulin-producing beta-cells promises an efficient approach to achieving euglycemia in IDDM patients. However, immune rejection limits long-term graft survival. Induction of islet-specific tolerance without the requirement for chronic immunosuppression would, therefore, offer a major advancement. T cells, directed at unique pancreatic beta-cell antigens, play the most critical role in islet cell destruction and development of IDDM. In this grant application, we propose to induce apoptosis in autoreactive and alloreactive lymphocytes by IL-2R-targeted delivery of caspase-activating molecules to treat diabetes. Apoptosis is the most potent physiological cell death that regulates immune homeostasis and tolerance to self-antigens. Dysregulation of the apoptotic process leads to autoimmunity, including diabetes. Caspases are the most important molecules that regulate the apoptotic machinery. Caspases are synthesized as inactive proenzymes in all the cells of the body. The activation of one caspase molecule is sufficient to commit the cell to death since it can activate the other caspases (>13 in mammals) by transcatalysis. Once activated, caspases override all the regulatory processes and commit the cell to death. Inasmuch as activated autoreactive and alloreactive T cells express high affinity IL-2R, we hypothesize that these cells can be targeted for apoptosis using IL-2R chimeric with caspase-activating proteins. Physical elimination of auto and alloreactive T cells is expected to result in the prevention of disease in prediabetic and treatment in diabetic animals transplanted with islet allografts. This approach may have direct clinical application as chimeric proteins can easily be generated in large quantities using recombinant DNA technology. The Specific Aims of this proposal include: 1) constructing recombinant genes encoding IL-2 chimeric with caspase-activating molecules and assessing chimeric proteins for specific apoptosis of autoreactive and alloreactive lymphocytes in vitro; 2) testing chimeric proteins in vivo for the prevention of diabetes in prediabetic NOD mice; 3) investigating the ability of chimeric proteins to induce tolerance to autoantigens and alloantigens for the prevention of islet allograft rejection and recurrence of autoimmunity.

描述（由申请人提供）：这笔赠款的主要目的是 特别诱导自动和同种异体淋巴细胞的细胞凋亡 激活内源性胱天蛋白酶。物理消除这些淋巴细胞 预计NOD小鼠将消除糖尿病前期的自身免疫过程 动物并防止患病动物的同种异体移植排斥，导致 预防和治疗糖尿病。 I型胰岛素依赖性糖尿病（IDDM）是一种慢性自身免疫性 影响全球大量人的疾病。 进步 理解IDDM发病机理的分子和细胞基础 医学实践的改善导致有效的治疗 预防IDDM的方法。尽管医疗疗法进步， IDDM仍然是长期发病率和死亡率的主要原因。是 需要移植的肾衰竭的最常见原因。 移植整个胰腺或纯化的胰岛 产生胰岛素的β细胞有望实现一种有效的方法 IDDM患者中的尤格西血症。但是，免疫排斥限制了长期移植 生存。诱导胰岛特异性公差无需 因此，慢性免疫抑制将提供重大进步。 T细胞针对独特的胰腺β细胞抗原，发挥最大的作用 IDDM的胰岛细胞破坏和发展中的关键作用。在这笔赠款中 应用，我们建议诱导自动反应性和同种异体反应性凋亡 通过IL-2R靶向caspase激活分子递送至淋巴细胞 治疗糖尿病。凋亡是最有效的生理细胞死亡 调节免疫稳态和对自我抗原的耐受性。失调的 凋亡过程导致自身免疫性，包括糖尿病。 caspase是 调节凋亡机制的最重要分子。 caspase 在人体所有细胞中被合成为无活性性酶。这 一个caspase分子的激活足以使细胞死亡 由于它可以通过经催化激活其他胱天蛋白酶（> 13个哺乳动物）。 激活后，caspases覆盖了所有监管过程，并提交 细胞死亡。 激活自动反应性和同种异体T细胞 表达高亲和力IL-2R，我们假设可以针对这些细胞 使用IL-2R嵌合与caspase激活蛋白的凋亡。身体的 消除自动和同种异体T细胞有望导致 预防糖尿病动物前糖尿病和治疗的疾病 用同种异体移植物移植。这种方法可能具有直接的临床 可以大量生成作为嵌合蛋白的应用 使用重组DNA技术。 该提案的具体目的包括：1）构建重组基因 用caspase激活分子编码IL-2嵌合并评估 嵌合蛋白，用于自动反应性和同种异体反应性特异性凋亡 体外淋巴细胞； 2）在体内测试嵌合蛋白预防 糖尿病前小鼠的糖尿病； 3）研究嵌合的能力 蛋白质诱导对自身抗原和同种抗原的耐受性 预防同种异体移植排斥和自身免疫的复发。

项目成果

Chronic cardiac allograft rejection in a rat model disparate for one single class I MHC molecule is associated with indirect recognition by CD4(+) T cells.

• DOI: 10.1016/s0966-3274(03)00004-2
• 发表时间: 2003-04
• 期刊: Transplant immunology
• 影响因子: 1.5
• 作者: H. Shirwan;A. Mhoyan;E. Yolcu;X. Que;S. Ibrahim

ProtEx: a novel technology to display exogenous proteins on the cell surface for immunomodulation.

ProtEx：一种在细胞表面展示外源蛋白以进行免疫调节的新技术。

• DOI: 10.1196/annals.1352.036
• 发表时间: 2005
• 期刊: Annals of the New York Academy of Sciences
• 影响因子: 5.2
• 作者: Singh,NarendraP;Yolcu,EsmaS;Askenasy,Nadir;Shirwan,Haval
• 通讯作者: Shirwan,Haval

Predominant expression of Th2 cytokines and interferon-gamma in xenogeneic cardiac grafts undergoing acute vascular rejection.

经历急性血管排斥反应的异种心脏移植物中 Th2 细胞因子和干扰素-γ 的主要表达。

• DOI: 10.1097/01.tp.0000052594.83318.68
• 发表时间: 2003
• 期刊: Transplantation.
• 作者: Singh,NarendraP;Guo,Luping;Mhoyan,Anna;Shirwan,Haval
• 通讯作者: Shirwan,Haval

Prophylactic fenbendazole therapy does not affect the incidence and onset of type 1 diabetes in non-obese diabetic mice.

预防性芬苯达唑治疗不会影响非肥胖糖尿病小鼠 1 型糖尿病的发病率和发作。

• DOI: 10.1093/intimm/dxh385
• 发表时间: 2006
• 期刊: International immunology.
• 作者: Franke,DeannaDH;Shirwan,Haval
• 通讯作者: Shirwan,Haval

A novel approach to cancer immunotherapy: tumor cells decorated with CD80 generate effective antitumor immunity.

• 发表时间: 2003-07
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: Narendra P. Singh;E. Yolcu;D. Taylor;Ç. Gerçel-Taylor;D. Metzinger;Stephen K Dreisbach;H. Shirwan