Unraveling the tolerogenic potential of lymph node fibroblastic reticular networks in autoimmune diabetes

揭示自身免疫性糖尿病中淋巴结成纤维细胞网状网络的耐受潜力

基本信息

批准号：
10624501
负责人：
Remi J Creusot
金额：
$ 17.83万
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：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10624501
关键词：
3-Dimensional Address Affect Antigen Targeting Antigen-Presenting Cells Antigens Autoantigens Autoimmune Diabetes Autoimmune Diseases Autoimmunity B-Lymphocytes Biomedical Engineering CCL21 gene CD8-Positive T-Lymphocytes Cells Child Chronic Cytoskeleton Data Defect Development Diabetes prevention Disease Engineering Frequencies Genetic Engineering Human Hybrids Immune Immune Tolerance Immune response Immunology Immunosuppression Impairment In Vitro Inbred NOD Mice Incidence Insulin Insulin-Dependent Diabetes Mellitus Kidney Knock-out Knowledge Lead Maintenance Modeling Mus Organ Organ Donor Pancreas Peptides Peripheral Phenotype Professional Role Property Regimen Regulation Regulatory T-Lymphocyte Reticular Cell Self Tolerance Site Societies Specificity Stromal Cells Supporting Cell T cell anergy T-Cell Proliferation T-Cell Receptor T-Lymphocyte Testing Therapeutic Thymus Gland Tissue Engineering Tissues Transgenic Mice Transgenic Organisms Transplantation Work adaptive immunity anergy antigen-specific T cells autoreactive B cell autoreactive T cell autoreactivity base capsule cellular engineering central tolerance clinical translation clinically translatable conventional therapy cytotoxicity engineered T cells immunoregulation implantation improved in vivo innovation islet loss of function lymph nodes mouse model novel strategies novel therapeutics overexpression peripheral tolerance prevent recruit subcutaneous therapeutic evaluation

项目摘要

Project Summary: Impaired central and peripheral tolerance cause type-1 diabetes (T1D); b-cell antigen-reactive T cells escape negative selection in the thymus and regulation/deletion in lymph nodes. The role of professional antigen- presenting cells and regulatory T cells in maintaining peripheral tolerance has been demonstrated and is currently being evaluated for therapy of autoimmune diseases, including T1D. Conversely, non-professional antigen-presenting cells like lymph node stromal cells are more stable tolerogenic cells but their potential for therapeutic tolerance induction in T1D has not been tested yet. Fibroblastic reticular cells (FRCs) are a subtype of lymph node stromal cells that support and remodel the lymph node and the integrity of FRC reticula is critical for adaptive immunity. In transgenic mice, expression and presentation of model antigens by FRCs to specific T cells induced their proliferation followed by deletion and absence of MHC-II on lymph node stromal cells was associated with progressive development of autoimmunity. Thus, FRC engineering for peripheral overexpression of tissue-specific antigens, including those not expressed in the thymus, may be exploited for tolerance induction in the therapeutic treatment of autoimmunity. We showed that expression of insulin, a major b-cell antigen, the relative FRC frequency and the FRC reticular remodeling properties are decreased in T1D. Thus, to exploit the capability of FRCs to promote antigen-specific tolerance for therapeutic treatment of T1D, FRCs need to be engineered to (i) increase their frequency by therapeutic transplantation and (ii) overexpress disease-relevant antigens. We recently developed genetic engineering approaches to overexpress T1D-relevant antigens, including hybrid peptides, in FRCs and tissue engineering approaches to fabricate three-dimensional FRC reticula that recapitulate FRC organization in the lymph node paracortex. Engineered FRC reticula overexpressing b-cell antigens and lacking expression of costimulatory molecules engraft in mice to recruit antigen-specific T cells and in vitro, engineered FRCs promote engagement of specific T cells, anergy, and regulatory T cell expansion, which could promote antigen-specific tolerance in vivo. Thus, in aim 1.1, we will test the therapeutic application of our engineered FRC reticula for tolerance induction and mechanisms in vitro and in vivo in a murine T1D model with selected disease-relevant antigens. For clinical translation, in aim 1.2, we will evaluate whether reticula engineered with b-cell antigen-expressing FRCs from human lymph nodes can engage human T cells engineered for antigen specificity, which is important for clinical translation of our approach. Peripheral expression and presentation of tissue-specific antigens by lymph node stromal cells to autoreactive T cells escaping thymic deletion in lymph nodes could contribute to peripheral tolerance. In aim 2, we will test the contribution of b-cell antigen expression and presentation by lymph node FRCs on tolerization of b-cell antigen autoreactive CD8 T cells in a mouse model of T1D.

项目摘要：中枢和周围耐受性受损导致1型糖尿病（T1D）； B细胞抗原反应性T细胞逸出淋巴结中胸腺和调节/缺失的负选择。专业抗原的作用已经证明了细胞和调节性T细胞在维持周围耐受性中目前正在评估包括T1D在内的自身免疫性疾病的治疗。相反，非专业抗原呈递细胞（如淋巴结基质细胞）是更稳定的耐受性细胞，但其潜力是 T1D中的治疗耐受性诱导尚未测试。成纤维细胞网状细胞（FRC）是支持和重塑淋巴的淋巴结基质细胞的亚型节点和FRC网状的完整性对于适应性免疫至关重要。在转基因小鼠中，表达和 FRC对特定T细胞的模型抗原呈现诱导其增殖，然后缺失和淋巴结基质细胞上的MHC-II缺乏与自身免疫的进行性发展有关。因此，FRC工程用于组织特异性抗原的周围过表达，包括未表达的抗原在胸腺中，可以利用自身免疫治疗的治疗治疗中耐受性。我们展示了胰岛素的表达，一种主要的B细胞抗原，相对FRC频率和FRC网状重塑 T1D的性质降低。因此，利用FRC促进抗原特异性公差的能力对于T1D的治疗治疗，需要对FRC进行设计以（i）通过治疗增加其频率移植和（ii）与疾病相关的抗原。我们最近开发了过表达T1D相关抗原的基因工程方法，包括混合动力肽，在FRC和组织工程方法中，以制造三维FRC网状在淋巴结旁胶质中概括FRC组织。工程的FRC Enticula过表达B细胞抗原和缺乏共刺激分子的表达，植入小鼠中，以募集抗原特异性T细胞和在体外，设计的FRC促进特定T细胞的参与，厌食和调节性T细胞扩展，这可以在体内促进抗原特异性耐受性。因此，在AIM 1.1中，我们将测试治疗应用我们工程的FRC网状，用于体外耐受性诱导和机制，在鼠T1D中的体内与选定疾病相关的抗原模型。对于临床翻译，在AIM 1.2中，我们将评估是否由人淋巴结的B细胞表达FRC设计的网状抗原可以吸引人T细胞针对抗原特异性设计，这对于我们方法的临床翻译很重要。外围淋巴结基质细胞对自身反应性T细胞的组织特异性抗原表达和表达在淋巴结中逃避胸腺缺失可能有助于周围耐受性。在AIM 2中，我们将测试 B细胞抗原表达和淋巴结FRC表现对B细胞抗原的耐受的贡献 T1D小鼠模型中的自动反应性CD8 T细胞。