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。减少了 增殖，并增加免疫介导的β细胞凋亡，胰岛素分泌细胞变得不足以 充分调节身体血糖。T1D诊断后不久β细胞完全被诊断为 而Permanent Lost受到了最近的研究的挑战，研究表明，即使是长期患有 糖尿病残留的β细胞被保留，这从胰腺中胰岛素阳性的胰岛的存在来证明。 并通过增加混合餐耐力试验后内源性胰岛素的产生。因此，T1糖尿病可以 被认为是一种慢性疾病，在β细胞自身免疫破坏、再生、 以及对细胞凋亡的抵抗。在临床上，这种平衡明显倾向于β细胞的进行性丧失 然而，从理论上讲，通过减少损失来正常化β细胞的稳态是可能的， 恢复功能，促进残留β细胞的再生。 在理解β细胞生物学和核糖核酸治疗领域的最新进展为 在β细胞中选择性地对控制细胞的那些基因进行靶向操作的前所未有的机会 增殖、凋亡和自身免疫抵抗。 我们认为，针对胰岛细胞的RNA适配子可能是特异性递送 治疗性RNA对内源性和移植物基因表达的正负调节作用 β细胞。 RNA适配子是一种小的、抗核糖核酸酶的非免疫原性寡核苷酸，可以深入渗透 进入组织，并凭借其高亲和力和三维结构识别他们的目标 专一性。这些人工合成的抗体都是化学合成的，可以很容易地与小分子结合 干扰或小的激活RNA(siRNA或Sarna)，可以被选择来穿透细胞质 在目标绑定时。 我们最近通过一种非监督选择方法，生物信息学分析和经验 验证，两个单克隆RNA适配子能够在体内优先识别人β细胞。 长期目标是有益地调节β细胞的动态平衡和防御自身免疫 体内，我们决定将这些适体与能够上调PDL1和XIAP(允许β细胞)的小RNA偶联 分别对抗免疫攻击和抑制细胞凋亡)，下调p57Kip2(To 促进β细胞增殖)。使用未分离的人胰岛的初步体外数据表明， 由此产生的治疗性RNA/胰岛特异性适配子嵌合体有效地调节靶标的表达 基因。 有了这个提议，我们将检验适体-嵌合体可以在体内介导基因调控的假设 在人类胰岛细胞中。这应该允许通过防止β细胞的丢失来改变它们的功能和动态平衡，通过 增加了它们的增殖，并提供了免受自身免疫的保护。这种创新的方法可能会 允许开发基于体内靶向调节β的T1D新治疗策略 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