Reversing HIV T cell dysfunction by aptamer targeting of therapeutic siRNAs

通过适配体靶向治疗性 siRNA 逆转 HIV T 细胞功能障碍

基本信息

批准号：
9004624
负责人：
Eli Gilboa
金额：
$ 75.98万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9004624
关键词：
Acquired Immunodeficiency Syndrome Affinity Antibodies Antiviral Agents CD8B1 gene Cancer Patient Cell Culture Techniques Cells Chronic Development Disease Progression Drug abuse Engineering Exhibits Functional disorder Goals HIV HIV Infections Health Human Immune Immune System Diseases Immune response Immune system Immunity Immunotherapy In Vitro Infection Lead Ligand Binding Ligands Macaca Malignant Neoplasms Mediator of activation protein Modality Modeling Monoclonal Antibodies Mus Nucleic Acids Oligonucleotides PDCD1LG1 gene Patients Pharmaceutical Preparations RNA Interference Research SIV Severities Small Interfering RNA Specificity T-Lymphocyte Testing Therapeutic Agents aptamer arm base cost cost effective exhaust exhaustion immune function immunogenicity inhibitor/antagonist novel programs receptor reconstitution synergism targeted treatment therapeutic target

项目摘要

DESCRIPTION: HIV specific CD8+ T cells, a pivotal arm of the antiviral immune response, exhibit progressive dysfunction, known as "exhaustion". Severity of immune exhaustion correlates with disease progression, and is especially prevalent among drug abuse patients who often delay treatments that control HIV replication. PD-1, an immune inhibitory receptor and mediator of exhaustion in the setting of chronic infections and cancer, is upregulated on exhausted HIV-specific CD8+ T cells and in vitro blockade of its ligand with anti-PD- L1 antibodies restores, albeit partially, their immune functions. Given that exhausted T cells express multiple immune inhibitory receptors, the synergism of co-blockade of two receptors, and the limited effect of PD-L1 antibodies in vitro, strongly argues that PD-1 blockade alone will not be sufficient to rescue the full functionality of exhausted HIV specific CD8+ T cells. With the aim of developing a clinically feasible, cost-effective, and broadly applicable, treatment to reverse immune dysfunction in HIV-infected patients, our goal is to develop a single, chemically synthesized, nucleic acid based agent that can block the action of multiple inhibitory receptors in the same cell, and can be efficiently and specifically delivered to the exhausted T cells of the patient. The proposed approach is to inhibit downstream mediators of exhaustion like Blimp-1, Eomes, VHL or Sprouty-2, using RNAi in the form of siRNAs that are targeted to exhausted HIV-specific CD8+ T cells by conjugation to an oligonucleotide aptamer ligand that binds to receptors expressed on exhausted T cells, like PD-1, LAG3, orTim-3. The chemically synthesized aptamers, a novel platform for ligands with engineered specificity, offer significant advantages over the monoclonal antibody platform in terms of manufacture, cost, and reduced immunogenicity. Our lab has pioneered the use of aptamers as therapeutic agents and targeting ligands to modulate immunity in the setting of cancer, and have recently demonstrated the remarkable efficiency of aptamer-targeted siRNA delivery to circulating CD8+ T cells in mice. The central hypothesis of this proposal is that PD-1, LAG3, or Tim-3 aptamer targeted siRNA inhibition of mediators of exhaustion like Blimp- 1, Eomes, VHL or Sprout-2 will reverse the dysfunction of HIV-specific CD8+ T cells to an extent that is superior to that of blockade with monoclonal antibodies targeting inhibitory receptors like PD-1. The research plan is to generate high affinity aptamers to the aforementioned inhibitory receptors, construct aptamer-siRNA conjugates, and assess their ability, in comparison to blockade with antibodies, to restore immune functions in (i) Exhausted T cell cultured from HIV infected patients, (ii) HIV infected mice that are fully reconstituted with a human immune system, and (iii) an SIV/macaque model. This program will have developed a cost-effective and broadly applicable lead compound to test in patients chronically infected with HIV.

描述：艾滋病毒特异性CD8+ T细胞，抗病毒免疫响应的关键臂，暴露的进行性渐进式功能障碍，称为“精疲力尽”。免疫疲劳的严重程度与疾病进展相关，并且在滥用药物滥用患者中尤为普遍，这些患者通常会延迟控制HIV复制的治疗。 PD-1是一种在慢性感染和癌症中的免疫抑制受体和疲劳的介体，对耗尽的HIV特异性CD8+ T细胞进行了更新，并在其体外用抗PD-L1抗体对其配体进行了体外阻断，但部分恢复了其免疫功能。鉴于耗尽的T细胞表达多种免疫抑制受体，因此在体外的两个受体的共阻形成的协同作用以及PD-L1抗体在体外的有限作用，强烈认为，仅PD-1阻断将不足以挽救耗尽的HIV HIV特异性CD8+ T细胞的全部功能。与目的是开发临床上可行，成本效益且广泛适用的治疗方法，以逆转HIV感染患者的免疫功能障碍，我们的目标是开发一种单一的，化学合成的，核酸的剂，可以阻止多个抑制性受体在多个抑制性受体中的作用提出的方法是使用RNAi抑制疲劳的下游介体，例如Blimp-1，Eomes，VHL或Sproty-2，以siRNA的形式抑制siRNA的形式，这些siRNA旨在通过与寡核苷酸置换液结合与少量核苷酸置换液结合与受体的受体结合到耗尽的T细胞上，lag3细胞，lag3细胞，lag3细胞，lag3 cyt 3，laiv-nim lag3 to pd-pd-1，lneim-laivecific cd8+ t细胞。化学合成的适体是一个具有工程特异性配体的新型配体平台，就制造，成本和降低的免疫原性而言，比单克隆抗体平台具有显着优势。我们的实验室率先使用适体作为治疗剂，并靶向配体在癌症的情况下调节免疫史，并最近证明了适体靶向的siRNA递送以循环的CD8+ T细胞在小鼠中循环的效率。该提案的中心假设是，PD-1，LAG3或TIM-3 APATMER的靶向siRNA抑制替代者（如Blimp-1，Eomes，eomes，vhl或sprout-2）的介体抑制siRNA，将逆转HIV特异性CD8+ T细胞的功能障碍，而与单核抗生素相比，与单核抗生素相比，它比单核受体的受体更高。 The research plan is to generate high affinity aptamers to the prioritized inhibitory receptors, construct atamer-siRNA conjugates, and assess their ability, in comparison to blockade with antibodies, to restore immuno functions in (i) Exhausted T cell cultured from HIV infected patients, (ii) HIV infected mice that are fully reconstituted with a human immuno system, and (iii) an SIV/macaque model.该计划将开发一种具有成本效益且广泛适用的铅化合物，以在患有艾滋病毒的长期感染患者中进行测试。