Gene-engineered dendritic cell therapy for diabetics

描述（由申请人提供）： 1型糖尿病是一种自身免疫性疾病，其发病机制在于胰腺中胰岛中产生胰岛素的β细胞的选择性破坏。目前的胰岛素替代治疗策略在严格控制血糖方面并不完全有效。虽然完整的朗格汉斯胰岛移植有可能恢复生理血糖控制，但终身免疫抑制干预的要求带来了使胰岛移植功能障碍的巨大风险。然而，这些策略可能会导致一系列其他问题，包括肾衰竭和恶性肿瘤风险。这项合作应用结合了匹兹堡儿童医院糖尿病诊所、匹兹堡大学糖尿病研究所和癌症研究所的努力，其目的是扩展和调整我们最初成功的基于树突状细胞的方法，以延长 NOD 小鼠糖尿病的发病时间。我们将向新发糖尿病 NOD 小鼠注射基因工程 DC 来操纵免疫系统，目的是保留残留的 β 细胞质量，以维持血糖降低甚至恢复正常血糖。一旦我们确认了这一点，我们建议在新发糖尿病人类志愿者中启动一项安全性试验。在这种方法中，志愿者将被注射一次基因工程 DC，并通过不同的补充标准评估安全性。基因工程 DC 将由外周血单核细胞离体繁殖的自体 DC 组成，并用转录因子 NF-kB (NF-kB ODN) 的短双链寡脱氧核糖核苷酸诱饵处理。在类似的方法中，将使用靶向 CD40、CD80 和 CD86 初级转录物的反义寡核苷酸代替 NF-kB 诱饵。我们将此应用程序分为两个部分。第一项将主要在 NOD 小鼠中进行研究，以证明原理，并最终申请在人类糖尿病志愿者中进行有限的安全性试验。第二项将把安全性试验扩展到更多的糖尿病患者群体，并最终对我们的方法保留残余 β 细胞质量和功能的潜力进行初步检查，通过刺激的 C 肽水平和血红蛋白 Ale 作为替代标记来评估。如果结果令人鼓舞，则可以在更多人群中尝试这些方法，以降低胰岛素需求、减少剂量，甚至逐渐减少胰岛素用量。最终结果可能包括降低新诊断糖尿病患者的胰岛素替代治疗强度，甚至完全停止胰岛素替代治疗，或许还有一种预防糖尿病的方法。

DESCRIPTION (provided by applicant): Type 1 diabetes mellitus is an autoimmune disease whose etiopathogenesis lies in the selective destruction of the insulin-producing beta cells of the islets of Langerhans in the pancreas. The current insulin replacement therapy strategies are not fully effective at recapitulating tight glucose control. While transplantation of intact islets of Langerhans offers the potential to restore physiologic glycemic control, the requirement for life-long immunosuppressive interventions carries with it significant risks of rendering the islet transplants dysfunctional. However, these strategies can lead to an array of other problems including kidney failure and a risk of malignancy. The objective of this collaborative application combining the efforts of the Diabetic Clinic of the Children's Hospital of Pittsburgh, The Diabetes Institute and the Cancer Institute of the University of Pittsburgh is to extend and adapt our initial successful dendritic cell-based approach of prolonging the onset time of diabetes in the NOD mouse. We will inject new-onset diabetic NOD mice with gene engineered DC to manipulate the immune system with the objective of preserving residual beta cell mass that can sustain reduced glycemia or even a return to normoglycemia. Once we confirm this, we propose to initiate a safety trial in new-onset diabetic human volunteers. In this approach, volunteers will be given a single injection of gene-engineered DC and safety will be assessed by different and complementary criteria. The gene-engineered DC will consist of autologous DC propagated ex vivo from peripheral blood mononuclear cells and treated with short, double-stranded oligodeoxyribonucleotide decoys for the transcription factor NF-kB (NF-kB ODN). In a similar approach, antisense oligonucleotides targeted to the CD40, CD80 and CD86 primary transcripts will be used in place of the NF-kB decoys. We divide this application into two sections. The first will deal with studies primarily in the NOD mouse to demonstrate proof-of-principle and will culminate in a application for a limited safety trial in human diabetic volunteers. The second will extend the safety trial into a larger population of diabetics and will culminate in a preliminary examination of the potential of our approach to preserve residual beta cell mass and function as assessed by stimulated C-peptide level and hemoglobin Ale as a surrogate marker. If the results are encouraging, these approaches could be attempted in a larger human population with an objective of lowered insulin requirements, decreased dosage and perhaps a gradual tapering off of insulin. The ultimate result could include less intensive or even a complete cessation of insulin replacement therapy in newly-diagnosed diabetics as well as, perhaps, a means of preventing diabetes.