Aptamer chimeras for the in vivo modulation of beta cell mass and immunogenicity

用于体内调节 β 细胞质量和免疫原性的适体嵌合体

• 批准号: 9458544
• 负责人: Midhat H Abdulreda
• 金额: $ 196.46万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-20 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9458544
• 关键词: Adenovirus Vector; Adenoviruses; Adult; Affinity; Apoptosis; Apoptosis Inhibitor; Apoptotic; Autoimmune Process; Autoimmunity; B Cell Proliferation; BIRC4 gene; Back; Beta Cell; Binding; Bioinformatics; Blindness; C-Peptide; CASP7 gene; CASP9 gene; Cadaver; Cell Count; Cell Death; Cell Proliferation; Cell physiology; Cells; Cellular biology; Cessation of life; Chemicals; Chimera organism; Chronic; Chronic Disease; Clinical; Clinical Trials; Cytoplasm; Data; Development; Diabetes Mellitus; Diagnosis; Disease; Engraftment; Equilibrium; FDA approved; Face; Fatty acid glycerol esters; Functional disorder; Future; Gene Expression; Gene Targeting; Generations; Genes; Goals; Homeostasis; Human; Hyperglycemia; Hypoglycemia; Hypoxia; Iatrogenesis; Image; Immune; Immunity; Immunologics; Immunosuppression; In Vitro; Inbred NOD Mice; Injection of therapeutic agent; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans Transplantation; Kidney Failure; Left; Leukocytes; Life; Link; Mediating; Medical; Methods; Mus; Myocardial Infarction; Natural regeneration; Nerve; Oligonucleotides; PDCD1LG1 gene; Pancreas; Pathology; Patients; Persons; Pharmaceutical Preparations; Pregnancy Complications; Production; Proliferating; Property; RNA; Research; Residual state; Resistance; Ribonucleases; Site; Small Interfering RNA; Small RNA; Specificity; Stroke; Structure of beta Cell of islet; T-Lymphocyte; Testing; Therapeutic; Time; Tissues; Transplantation; Untranslated RNA; Up-Regulation; Validation; anterior chamber; aptamer; base; cancer therapy; cancer type; cytokine; cytotoxicity; diabetic; experimental study; eye chamber; fighting; functional loss; glucose metabolism; human disease; humanized mouse; immunogenicity; improved; in vivo; innovation; insulin secretion; islet; isoimmunity; mouse model; novel; novel strategies; novel therapeutics; polypeptide C; prevent; protective effect; synthetic antibodies; targeted treatment; therapeutic RNA; three dimensional structure; tool

Type 1 (T1D) diabetes results from the immune mediated loss of β cell mass and function. With a reduced proliferation, and an increase immune mediated β cell apoptosis, insulin producing cells become insufficient to adequately regulate body glycemia. The historical dogma for which soon after T1D diagnosis β cell were fully and permanently lost has been challenged by recent studies revealing that even patients with long-standing diabetes residual β cells are retained as demonstrated by the presence of insulin positive islets in the pancreas and by increase production of endogenous insulin after mixed-meal tolerance test. T1 diabetes can thus been seen as a chronic disease in which the balance exist between β cells autoimmune destruction, regeneration, and resistance to apoptosis. Clinically this balance is obviously tilted toward the progressive loss of β cell mass, however, it might be theoretically possible to normalize β cell homeostasis by reducing loss, recovering function, and enhancing regeneration of remnant β cells. Recent advancements in the understanding β cell biology and in the field of RNA therapeutics offer an unprecedented opportunity for a targeted manipulation selectively in β cells of those genes that control cell proliferation, apoptosis and resistance to autoimmunity. We propose that RNA aptamers specific for islet cells might be the tool for the specific delivery of therapeutic RNA able to positively and negatively modulate gene expression in endogenous and transplanted β cells. RNA aptamers are small, RNAse resistant, non-immunogenic oligonucleotide that can penetrate deeply into the tissue and recognize their target in virtue of their three dimensional structure with high affinity and specificity. These synthetic antibodies are chemically synthetized, can be easily conjugated with small interference or small activating RNAs (siRNA or saRNA) and can be selected to penetrate into the cytoplasm upon target binding. We have recently identified by an unsupervised selection method, bioinformatics analysis and empirical validation, two monoclonal RNA aptamers able to recognized preferentially human β cells in vivo. With the long term goal to beneficially modulate β cell homeostasis and defense from auto-immunity in vivo, we decided to conjugate these aptamer with small RNA able to upregulate PDL1 and Xiap (to allow β cell to fight back the immunologic attack and inhibit apoptosis respectively) and to down-regulate p57kip2 (to promote β cell proliferation). Preliminary in vitro data using non-dissociated human islets demonstrated that the resulting therapeutic-RNA/islet-specific aptamer chimera effectively modulate the expression of the target genes. With this proposal we will test the hypothesis that aptamer-chimeras can be mediated in vivo gene modulation in human islet cells. This should allow to modify β cell function and homeostasis by preventing their loss, by increasing their proliferation, and by providing protection from autoimmunity. This innovative approach may allow the development of novel therapeutic strategies for T1D based on the in vivo targeted modulation of β cell genes.

1 型 (T1D) 糖尿病是由免疫介导的 β 细胞质量和功能丧失引起的。随着减少 增殖和免疫介导的 β 细胞凋亡增加，产生胰岛素的细胞变得不足以 充分调节体内血糖。在 T1D 诊断后不久，β 细胞就完全被完全接受的历史教条 最近的研究表明，即使是长期患有这种疾病的患者，也面临着永久迷失的挑战 胰腺中胰岛素阳性胰岛的存在证明了糖尿病残余 β 细胞的保留 并通过混合膳食耐受试验后增加内源性胰岛素的产生。因此，T1 型糖尿病可以 被视为一种慢性疾病，β细胞自身免疫破坏、再生、 和对细胞凋亡的抵抗。临床上这种平衡明显倾向于β细胞的逐渐丧失 然而，理论上可能可以通过减少损失来使 β 细胞稳态正常化， 恢复功能，增强残余β细胞的再生。 β 细胞生物学和 RNA 治疗领域的最新进展为我们提供了一个新的视角。 对β细胞中控制细胞的基因进行选择性靶向操作的前所未有的机会 增殖、凋亡和自身免疫抵抗。 我们提出，胰岛细胞特异性的RNA适体可能是特异性递送胰岛细胞的工具。 治疗性RNA能够正向和负向调节内源性和移植物中的基因表达 β细胞。 RNA适体是小的、RNA酶抗性、非免疫原性寡核苷酸，可以深入渗透 进入组织并凭借其高亲和力的三维结构识别其目标 特异性。这些合成抗体是化学合成的，可以很容易地与小分子缀合 干扰或小激活 RNA（siRNA 或 saRNA），可以选择渗透到细胞质中 目标结合后。 我们最近通过无监督选择方法、生物信息学分析和实证研究确定了 经验证，两种单克隆RNA适体能够在体内优先识别人β细胞。 长期目标是有益地调节 β 细胞稳态和防御自身免疫 在体内，我们决定将这些适体与能够上调 PDL1 和 Xiap 的小 RNA 结合（以允许 β 细胞 分别抵抗免疫攻击和抑制细胞凋亡）和下调 p57kip2（以 促进β细胞增殖）。使用非分离人类胰岛的初步体外数据表明， 由此产生的治疗性 RNA/胰岛特异性适体嵌合体有效调节靶标的表达 基因。 通过这个提议，我们将测试适体嵌合体可以介导体内基因调节的假设 在人类胰岛细胞中。这应该可以通过防止β细胞损失来改变β细胞功能和体内平衡， 增加它们的增殖，并提供针对自身免疫的保护。这种创新方法可能 允许基于β的体内靶向调节开发新的T1D治疗策略 细胞基因。

项目成果

RNA aptamers specific for transmembrane p24 trafficking protein 6 and Clusterin for the targeted delivery of imaging reagents and RNA therapeutics to human β cells.

• DOI: 10.1038/s41467-022-29377-3
• 发表时间: 2022-04-05
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Van Simaeys D;De La Fuente A;Zilio S;Zoso A;Kuznetsova V;Alcazar O;Buchwald P;Grilli A;Caroli J;Bicciato S;Serafini P
• 通讯作者: Serafini P

Integrated Metabolomics and Proteomics Analyses in the Local Milieu of Islet Allografts in Rejection versus Tolerance.

• DOI: 10.3390/ijms22168754
• 发表时间: 2021-08-15
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Hernandez LF;Betancourt LR;Nakayasu ES;Ansong C;Ceballos GA;Paredes D;Abdulreda MH
• 通讯作者: Abdulreda MH