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，其发病机理在于胰腺中胰岛的产生胰岛素的β细胞的选择性破坏。目前的胰岛素替代治疗策略在概括严格的血糖控制方面并不完全有效。虽然移植完整的胰岛提供了恢复生理血糖控制的可能性，但终身免疫抑制干预的要求带来了使胰岛移植物功能障碍的重大风险。然而，这些策略可能导致一系列其他问题，包括肾衰竭和恶性肿瘤的风险。这项合作应用的目的是结合匹兹堡儿童医院糖尿病诊所，糖尿病研究所和匹兹堡大学癌症研究所的努力，扩展和调整我们最初成功的基于树突状细胞的方法，延长NOD小鼠糖尿病的发病时间。我们将用基因工程DC注射新发糖尿病NOD小鼠，以操纵免疫系统，目的是保留残留的β细胞群，以维持降低的血糖，甚至恢复正常血糖。一旦我们证实了这一点，我们建议在新发糖尿病志愿者中启动一项安全性试验。在这种方法中，志愿者将接受单次基因工程DC注射，并通过不同的互补标准评估安全性。基因工程化的DC将由从外周血单核细胞离体繁殖的自体DC组成，并用转录因子NF-κ B的短双链寡脱氧核糖核苷酸诱饵（NF-κ B ODN）处理。在类似的方法中，将使用靶向CD 40、CD 80和CD 86初级转录物的反义寡核苷酸代替NF-κ B诱饵。我们将此应用程序分为两个部分。第一个将主要在NOD小鼠中进行研究，以证明原理性证据，并最终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大的糖尿病患者群体，并最终初步检查我们的方法保留残余β细胞质量和功能的潜力，如通过刺激的C肽水平和血红蛋白Ale作为替代标志物所评估的。如果结果令人鼓舞，这些方法可以在更大的人群中尝试，目的是降低胰岛素需求，减少剂量，也许逐渐减少胰岛素。最终的结果可能包括在新诊断的糖尿病患者中减少胰岛素替代疗法的强度，甚至完全停止胰岛素替代疗法，以及预防糖尿病的方法。

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.