Therapeutic efficacy of antisense oligonucleotides targeting PHLPP in preclinical models of diabetes

靶向PHLPP的反义寡核苷酸在糖尿病临床前模型中的治疗效果

• 批准号: 498542712
• 负责人: Dr. Amin Ardestani, Ph.D.
• 金额: --
• 依托单位: Zentrum für Biomolekulare Interaktionen (CBIB)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/498542712?language=en
• 关键词: Therapeutic; efficacy; antisense; oligonucleotides; targeting

Both type 1 diabetes (T1D) and type 2 diabetes (T2D), highly complex in their pathophysiology, finally result from pancreatic beta cell failure; hallmark and fundamental cause of diabetes. Underlying molecular mechanisms are still not fully characterized in order to provide beta cell specific therapies for a cure. The modulation of the survival of highly functional beta cells and/or enhancement of beta cell regenerative capacity therefore represent an attractive therapeutic approach for diabetes. Serine-threonine phosphatases (STPs) such as PHLPP1/2 (PH domain leucine-rich repeat protein phosphatase-1/2) regulate cell survival and serve as potential targets for drug development. Our understanding of the function and mechanism of PHLPPs in beta cell death regulation has important biological and clinical significance. We have identified that PHLPPs levels were highly elevated in metabolically stressed human and rodent diabetic β-cells. Genetic inhibition of PHLPPs markedly improved beta cell survival and function in experimental models of diabetes in vitro, in vivo and in islets from patients with T2D, presenting PHLPPs as promising targets for functional regenerative therapy of pancreatic beta cells in diabetes. With the aim of therapeutically inhibiting PHLPPs in a beta cell specific manner, this project will exploit a novel strategy of ligand-induced internalization of the glucagon-like peptide-1 receptor (GLP1R) to deliver PHLPP antisense oligonucleotides (PHLPP-ASOs) potently and specifically to pancreatic beta cells by conjugating them to a GLP1R peptide agonist. We will test GLP1-PHLPP-ASOs for their ability to restore beta cell survival and function and to normalize glycemia in vitro und in vivo. Also, we will identify the regulatory networks of direct or indirect PHLPP-driven transcriptional gene regulation in PHLPP-depleted β-cells at the bulk (whole islets) and single cell level (beta cells/endocrine cells) by using RNA-sequencing. This will help us to further characterize potential functional metabolic, proliferative and survival pathways with major focus on beta cell survival signaling such as mTOR, Hippo, AKT and OXPHOS. The purpose of this proposal is to move forward our initial highly promising results into translational research; to establish the previously uncharacterized PHLPP1/2 signaling pathway as a novel target for curative pharmacological intervention to restore functional pancreatic beta cells and normoglycemia in diabetes.

1型糖尿病（T1 D）和2型糖尿病（T2 D）两者在其病理生理学上高度复杂，最终由胰腺β细胞衰竭引起;胰腺β细胞衰竭是糖尿病的标志和根本原因。潜在的分子机制仍然没有完全表征，以提供用于治愈的β细胞特异性疗法。因此，调节高功能β细胞的存活和/或增强β细胞再生能力代表了糖尿病的有吸引力的治疗方法。丝氨酸-苏氨酸磷酸酶（STP）如PHLPP 1/2（PH结构域富含亮氨酸重复蛋白磷酸酶-1/2）调节细胞存活，并作为药物开发的潜在靶点。了解PHLPPs在β细胞死亡调控中的功能和机制具有重要的生物学和临床意义。 我们已经确定PHLPP水平在代谢应激的人和啮齿动物糖尿病β细胞中高度升高。PHLPP的遗传抑制在体外、体内和来自T2 D患者的胰岛中显著改善了糖尿病实验模型中的β细胞存活和功能，这表明PHLPP是糖尿病中胰腺β细胞的功能性再生治疗的有希望的靶点。为了以β细胞特异性方式治疗性抑制PHLPP，该项目将利用胰高血糖素样肽-1受体（GLP 1 R）的配体诱导内化的新策略，通过将PHLPP反义寡核苷酸（PHLPP-ASO）缀合至GLP 1 R肽激动剂，将其有效且特异性地递送至胰腺β细胞。我们将测试GLP 1-PHLPP-ASO在体外和体内恢复β细胞存活和功能以及使β细胞正常化的能力。此外，我们将通过使用RNA测序在大量（整个胰岛）和单细胞水平（β细胞/内分泌细胞）鉴定PHLPP耗尽的β细胞中直接或间接PHLPP驱动的转录基因调控的调控网络。这将有助于我们进一步表征潜在的功能性代谢、增殖和存活途径，主要关注β细胞存活信号传导，如mTOR、Hippo、AKT和OXPHOS。该提案的目的是将我们最初非常有希望的结果推进到转化研究中;建立以前未表征的PHLPP 1/2信号通路作为治疗性药理学干预的新靶点，以恢复糖尿病中功能性胰腺β细胞和正常胰岛素。