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：
10672991
负责人：
Remi J Creusot
金额：
$ 15.39万
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/10672991
关键词：
3-Dimensional Address Affect Antigen Presentation 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 Engraftment 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 Maintenance Modeling Mus Organ Organ Donor Pancreas Peptides Peripheral Phenotype Professional Role Proliferating Property Regimen Regulation Regulatory T-Lymphocyte Relaxation Reticular Cell Reticulum Self Tolerance Site Societies Specificity Stromal Cells Supporting Cell 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 capsule cellular engineering central tolerance clinical translation 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 reticulum cell 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.

项目概要：中枢和外周耐受性受损导致 1 型糖尿病 (T1D)； b 细胞抗原反应性 T 细胞逃逸胸腺的负选择和淋巴结的调节/删除。专业抗原的作用- 呈递细胞和调节性 T 细胞在维持外周耐受方面的作用已被证明并且正在研究目前正在评估其治疗自身免疫性疾病（包括 T1D）的效果。反之，非专业人士淋巴结基质细胞等抗原呈递细胞是更稳定的耐受性细胞，但它们具有潜在的耐受性。 T1D 的治疗耐受诱导尚未经过测试。成纤维细胞网状细胞 (FRC) 是淋巴结基质细胞的一种亚型，支持和重塑淋巴节点和 FRC 网状结构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 将模型抗原呈递给特定 T 细胞，诱导其增殖，然后删除和淋巴结基质细胞上 MHC-II 的缺失与自身免疫的进行性发展相关。因此，FRC工程用于组织特异性抗原的外周过度表达，包括那些不表达的抗原在胸腺中，可用于自身免疫治疗中的耐受诱导。我们展示了胰岛素（一种主要 B 细胞抗原）的表达、相对 FRC 频率和 FRC 网状重塑 T1D 的特性下降。因此，利用 FRC 促进抗原特异性耐受的能力对于 T1D 的治疗，需要对 FRC 进行改造以 (i) 通过治疗提高其频率移植和（ii）过度表达疾病相关抗原。我们最近开发了基因工程方法来过度表达 T1D 相关抗原，包括杂交肽，在 FRC 和组织工程方法中制造三维 FRC 网状结构概括一下副皮质淋巴结中的 FRC 组织。工程化 FRC 网状结构过度表达 b 细胞抗原和缺乏共刺激分子的表达移植到小鼠体内以招募抗原特异性T细胞和在体外，工程化 FRC 促进特定 T 细胞的参与、无反应性和调节性 T 细胞扩增，可以促进体内抗原特异性耐受。因此，在目标 1.1 中，我们将测试治疗应用我们工程化的 FRC 网状结构在小鼠 T1D 中的体外和体内耐受诱导和机制具有选定的疾病相关抗原的模型。对于临床转化，在目标 1.2 中，我们将评估是否用来自人类淋巴结的表达 b 细胞抗原的 FRC 改造的网状结构可以与人类 T 细胞结合专为抗原特异性而设计，这对于我们方法的临床转化非常重要。周边淋巴结基质细胞向自身反应性 T 细胞表达和呈递组织特异性抗原逃避淋巴结中的胸腺缺失可能有助于外周耐受。在目标 2 中，我们将测试淋巴结 FRC 的 b 细胞抗原表达和呈递对 b 细胞抗原耐受的贡献 T1D 小鼠模型中的自身反应性 CD8 T 细胞。