In Vivo Prevention of Murine GVHD

小鼠 GVHD 的体内预防

• 批准号: 10610863
• 负责人: Bruce R Blazar
• 金额: $ 58.97万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610863
• 关键词: Mus; Prevention; S-7; in vivo

Abstract Our overall goals are focus on the prevention and treatment of graft-vs-host disease (GVHD) via innate lymphoid type 2 cell (ILC2) anti-inflammatory and tissue reparative properties. We found that host ILC2s in are eliminated by total body irradiation {TBI} or chemotherapy and remain depleted for at >/=90 days. This finding is highly relevant as there is an inverse correlation between peripheral blood activated ILC2s and GVHD. As such. we sought to determine whether supplemental infusion of mature donor ILC2s could be used to prevent murine GVHD. We showed for the first time that donor ILC2s could prevent or partially treat GVHD in an amphiregulin (AREG) dependent process. Whether AREG or ILC2 direct contact with intestinal stem cells {ISC) supports small intestine epithelial cell repair in TBI treated mice or organoids is unknown. Additionally, third-party ILC2 infusion also significanUy reduced murine GVHD lethality. lmportanUy for translational purposes, we found ILC2s to be relatively steroid resistant. Peri-BMT {bone marrow transplant) IL-33 increased ST2/IL33R+ ILC2s at BMT day 0 and reduced GVHD. Ko mice had accelerated GVHD; IL-33 given pre-BMT prevented the full lLC2 loss. IL-33 ko recipients have hypo-proliferative epithelial cells, reduced ISCs and Paneth cells, and smaller crypt height and numbers. Ex vivo intestine organoid culture modeling revealed that IL-33 coordinated regeneration by inducing epidermal growth factor (EGF), significantly reduced by TBI. EGF restored ISC deficiency, uncovering a gut repair IL-33/EGF loop between ISCs and Paneth cells. Donor IL-13 ko ILC2s or host IL 13Ra ko mice had reduced GVHD. ILC2 IL 13 supports both ST2+ tuft cells and goblet cells. Tuft cells produce IL-25 driving ILC2 production and survival. TBI markedly reduced tuft cells for :!:38 days and ko recipients had a striking increase in GVHD. When given to wildtype mice exogenous IL-25 significantly reduced GVHD. Consequences of ko of tuft cells (and ILC2s) on donor T cell expansion, trafficking and function are unknown. The role of IL-25 and IL 17RB has not been examined. We will address the dynamics and interplay between ILC2, IL-33 and host intestinal cells (tuft cells, ISCs, Paneth cells) after TBI and during GVHD. Aim 1 will test the hypothesis that: Donor ILC2 repopulation fails due to destruction of ILC2 BM niche that supports ILC2s. In vitro pre-lLC2 differentiation, maturation and expansion ± proinflammatory cytokines and ILC2 supporting cytokines will be studied. GATA3-GFPhi pre-lLC2s/mature ILC2s transfer into lethally irradiated congenic BMT recipients will provide data on the differential ability to repopulate the BM. If the BM cannot support pre-lLC2s/lLC2s, we will study stem cell deficient mice. Aim 2 will test the hypothesis that: Pre-lLC2s/lLC2s and their secreted products have direct effects on intestinal cell subsets. Intestinal organoid cultures from wild-type and IL-33 ko mice with syngeneic Tregs and ILC2s will measure organoid size, number, and gene expression related to proliferation, cell cycle regulation, and specific epithelial lineage markers under homeostasis or after TBI. Anti-AREG mAbs or co-cultures with AREG ko lymphocytes will be characterized for promoting epithelial regeneration. Aim 3 will test the hypothesis that: Tuft cells are essential for ILC2 development and survival via an ILC2 release of IL-13 that stimulates tuft cells to release IL-25 that causes ILC2 proliferation (aim 3). Tuft cell and ILC2 ko hosts have accelerated GVHD. We hypothesize that IL-25 effects are due to direct stimulation of ILC2s or alternatively, with donor IL-17RB+ T cells. Significance. Studies in the R37 extension phase will provide fundamental information as to the mechanisms by which peri-BMT IL-33 diminish GVHD lethality via effects on ST2+ host ILC2s and regulatory T cells, both of which produce AREG, and the EGF/IL-33 loop that occurs between ISCs and Paneth cells resulting in small intestine repair. Further, the key role of host ILC2s in subduing GVHD, the nature of post-BMT ILC2 deficiency that occurs after pre-BMT conditioning regimens and predisposes patients to GVHD, and the essential requirement for tuft cells or their product, IL-25 will be elucidate. LasUy, critical insights will be gained as to the inability of pre-lLC2s generation, gut migration or differentiation. Translational Impact: The efficacy of donor and third-party ILC2 infusion in preventing and treating GVHD support our planned human ILC2 clinical trial, funded via other auspices, that will infuse "off-the-shelf' third party ILC2s to treat steroid refractory gut GVHD, that portends a particularly poor prognosis, in a 2- institutional study at the University of Minnesota and UNC-CH.

抽象的 我们的总体目标是通过先天淋巴预防和治疗移植物抗宿主病 (GVHD) 2 型细胞 (ILC2) 的抗炎和组织修复特性我们发现宿主 ILC2 被消除。 通过全身照射 {TBI} 或化疗，并在 >/=90 天时保持耗尽状态，这一发现非常重要。 相关，因为外周血激活的 ILC2 和 GVHD 之间存在负相关。 试图确定是否可以使用成熟供体 ILC2 的补充输注来预防小鼠 GVHD。我们首次证明供体 ILC2 可以预防或部分治疗双调蛋白中的 GVHD。 (AREG)依赖过程是否与肠干细胞(ISC)直接接触AREG或ILC2支持小。 此外，第三方 ILC2 输注对 TBI 治疗小鼠或类器官的肠上皮细胞修复作用尚不清楚。 我们发现 ILC2 还显着降低了小鼠 GVHD 致死率。 周围 BMT（骨髓移植）IL-33 在 BMT 日增加 ST2/IL33R+ ILC2。 0 和 GVHD 降低的 Ko 小鼠加速了 GVHD；BMT 前给予 IL-33 阻止了 IL-33 的完全丧失。 ko 受者的上皮细胞增殖不足、ISC 和潘氏细胞减少、隐窝高度较小 离体肠道类器官培养模型显示 IL-33 通过协调再生。 诱导表皮生长因子 (EGF) 因 TBI 显着减少而恢复了 ISC 缺陷，揭示了这一点。 ISC 和潘氏细胞之间的肠道修复 IL-33/EGF 环供体 IL-13 ko ILC2 或宿主 IL 13Ra ko 小鼠具有。 降低 GVHD。IL 13 支持 ST2+ 簇状细胞和杯状细胞产生驱动 ILC2 的 IL-25。 TBI 显着减少簇细胞，持续 38 天，而 KO 接受者则显着增加。 当给予野生型小鼠外源性IL-25时，GVHD的后果显着降低。 IL-25 和簇细胞（和 ILC2）对供体 T 细胞扩增、运输和功能的作用尚不清楚。 IL 17RB 尚未经过检查，我们将讨论 ILC2、IL-33 和之间的动态和相互作用。 TBI 后和 GVHD 期间的宿主肠细胞（簇细胞、ISC、潘氏细胞）。 目标 1 将检验以下假设： 由于 ILC2 BM 生态位遭到破坏，供体 ILC2 重新增殖失败 支持 ILC2 体外分化、成熟和扩增 ± 促炎性。 将研究细胞因子和 ILC2 支持细胞因子 GATA3-GFPhi pre-lLC2s/成熟 ILC2s 转移。 进入受致命辐射的同源 BMT 接受者将提供有关重新填充细胞的差异能力的数据 BM。如果 BM 不能支持 pre-lLC2s/lLC2s，我们将研究干细胞缺陷小鼠，目标 2 将测试。 假设：Pre-lLC2s/lLC2s 及其分泌产物对肠细胞亚群有直接影响。 来自野生型和具有同基因 Tregs 和 ILC2 的 IL-33 ko 小鼠的肠道类器官培养物将进行测量 类器官的大小、数量和与增殖、细胞周期调节和特异性相关的基因表达 稳态下或 TBI 后的上皮谱系标记物或与 AREG ko 共培养。 淋巴细胞将被表征为促进上皮再生。目标 3 将检验以下假设：簇绒。 细胞对于 ILC2 的发育和存活至关重要，ILC2 释放 IL-13，刺激簇细胞 释放引起ILC2增殖的IL-25（目标3）并且ILC2 ko宿主加速了GVHD。 IL-25 效应是由于 ILC2 的直接刺激或供体 IL-17RB+ T 的直接刺激所致 细胞。 意义。R37 延伸阶段的研究将提供有关机制的基本信息。 IL-3 通过对 BMT 周期间的 ST2+ 宿主 ILC2 和调节性 T 细胞的影响来降低 GVHD 致死率， 两者均产生 AREG，并且 ISC 和潘氏细胞之间发生的 EGF/IL-33 环导致 此外，宿主ILC2在抑制GVHD中的关键作用，BMT后ILC2的性质。 BMT 预处理方案后发生的缺陷使患者易患 GVHD，并且 簇细胞或其产品 IL-25 的基本要求将得到阐明，重要的见解将得到阐明。 获得了前 lLC2 生成、肠道迁移或分化的能力。 转化影响：供体和第三方 ILC2 输注在预防和治疗 GVHD 方面的功效 支持我们计划的人类 ILC2 临床试验，该试验由其他赞助资助，这将注入“现成的”第三种药物 一方 ILC2 治疗类固醇难治性肠道 GVHD，这预示着预后特别差，在 2- 明尼苏达大学和 UNC-CH 的机构研究。

项目成果

MicroRNA-155 Modulates Acute Graft-versus-Host Disease by Impacting T Cell Expansion, Migration, and Effector Function.

MicroRNA-155 通过影响 T 细胞扩增、迁移和效应器功能来调节急性移植物抗宿主病。

• 发表时间: 2018-06-15
• 作者: Zitzer, Nina C;Snyder, Katiri;Meng, Xiamoei;Taylor, Patricia A;Efebera, Yvonne A;Devine, Steven M;Blazar, Bruce R;Garzon, Ramiro;Ranganathan, Parvathi
• 通讯作者: Ranganathan, Parvathi

Dendritic Cell Expression of Retinal Aldehyde Dehydrogenase-2 Controls Graft-versus-Host Disease Lethality.

视网膜醛脱氢酶 2 的树突状细胞表达控制移植物抗宿主病致死率。

• 发表时间: 2019
• 作者: Thangavelu, Govindarajan;Lee, Yu;Loschi, Michael;Schaechter, K Melanie;Feser, Colby J;Koehn, Brent H;Nowak, Elizabeth C;Zeiser, Robert;Serody, Jonathan S;Murphy, William J;Munn, David H;Chambon, Pierre;Noelle, Randolph J;Blazar, Bruce R
• 通讯作者: Blazar, Bruce R

CD4 T cell sphingosine 1-phosphate receptor (S1PR)1 and S1PR4 and endothelial S1PR2 regulate afferent lymphatic migration.

CD4 T 细胞 1-磷酸鞘氨醇受体 (S1PR)1 和 S1PR4 以及内皮 S1PR2 调节传入淋巴管迁移。

• 发表时间: 2019
• 影响因子: 24.8
• 作者: Xiong, Yanbao;Piao, Wenji;Brinkman, C Colin;Li, Lushen;Kulinski, Joseph M;Olivera, Ana;Cartier, Andreane;Hla, Timothy;Hippen, Keli L;Blazar, Bruce R;Schwab, Susan R;Bromberg, Jonathan S
• 通讯作者: Bromberg, Jonathan S

Eomesodermin promotes the development of type 1 regulatory T (TR1) cells.

Eomesodermin 促进 1 型调节性 T (TR1) 细胞的发育。

• 发表时间: 2017-04-07
• 影响因子: 24.8
• 作者: Zhang, Ping;Lee, Jason S;Gartlan, Kate H;Schuster, Iona S;Comerford, Iain;Varelias, Antiopi;Ullah, Md Ashik;Vuckovic, Slavica;Koyama, Motoko;Kuns, Rachel D;Locke, Kelly R;Beckett, Kirrilee J;Olver, Stuart D;Samson, Luke D;Montes de Oca, Marc
• 通讯作者: Montes de Oca, Marc

IL-2 enhances ex vivo-expanded regulatory T-cell persistence after adoptive transfer.

IL-2 可增强过继转移后离体扩增的调节性 T 细胞的持久性。

• 发表时间: 2020-04-28
• 影响因子: 7.5
• 作者: Furlan, Scott N;Singh, Karnail;Lopez, Christina;Tkachev, Victor;Hunt, Daniel Joel;Hibbard, James;Betz, Kayla M;Blazar, Bruce R;Trapnell, Cole;Kean, Leslie S
• 通讯作者: Kean, Leslie S