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 我们的总体目标是专注于通过先天淋巴机构的预防和治疗移植物-VS宿主疾病（GVHD） 2型细胞（ILC2）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}或化学疗法，并保持在>/= 90天。这个发现很高 相关，因为外周血激活的ILC2和GVHD之间存在反相关。像这样。我们 试图确定是否可以使用补充成熟供体ILC2的补充输注来预防鼠 GVHD。我们首次表明捐助者ILC2可以预防或部分治疗两次gvhd （AREG）依赖过程。 AREG还是ILC2直接与肠道干细胞（ISC）直接接触都支持小 TBI处理的小鼠或类器官中的肠上皮细胞修复尚不清楚。此外，第三方ILC2输注 同样显着降低了鼠GVHD致死性。 lmportanuy用于翻译目的，我们发现ILC2是 BMT Day的Peri-BMT {骨髓移植）IL-33增加了ST2/IL33R+ ILC2S 0和减少GVHD。 KO小鼠加速了GVHD； IL-33给出的BMT防止了完整的LLC2损失。 IL-33 KO接受者的上皮细胞不变，ISC和Paneth细胞减少，并且加密高度较小 和数字。离体肠道器官培养建模表明，IL-33通过 诱导表皮生长因子（EGF），TBI显着降低。 EGF恢复了ISC缺乏症，发现 肠道修复IL-33/EGF循环在ISC和Paneth细胞之间。供体IL-13 KO ILC2S或宿主IL 13RA KO小鼠 减少GVHD。 ILC2 IL 13支持ST2+簇细胞和杯状细胞。 Tuft细胞产生IL-25驱动ILC2的IL-25 生产和生存。 TBI明显减少了簇式细胞：！：38天，KO接收者的罢工增加了 在GVHD中。当给予野生型小鼠时，外源IL-25显着降低了GVHD。 KO的后果 Tuft细胞（和ILC2）在供体T细胞扩张，运输和功能上尚不清楚。 IL-25和 IL 17RB尚未检查。我们将解决ILC2，IL-33和 TBI和GVHD之后，宿主肠细胞（簇细胞，ISC，Paneth细胞）。 AIM 1将检验以下假设：供体ILC2重生由于ILC2 BM生态位的破坏而失败 这支持ILC2。体外前LLC2分化，成熟和膨胀±促炎 细胞因子和支持细胞因子的ILC2将进行研究。 GATA3-GFPHI Pre-llc2s/成熟ILC2S转移 进入致命的辐照的先天性BMT接受者将提供有关重新填充差异能力的数据 BM。如果BM不能支持Pre-LLC2S/LLC2S，我们将研究干细胞缺乏小鼠。 AIM 2将测试 假设：前LLC2S/LLC2及其分泌产品对肠细胞亚群有直接影响。 来自野生型和IL-33 KO小鼠的肠道类器官培养物和ILC2的肠道培养物将测量 类器官大小，数量和基因表达与增殖，细胞周期调节和特定有关 稳态下或TBI之后的上皮谱系标记。抗肉体mab或与AREG KO的共同文化 淋巴细胞将用于促进上皮再生。 AIM 3将检验以下假设：簇 通过ILC2释放IL-13的ILC2发育和生存至关重要 释放导致ILC2增殖的IL-25（AIM 3）。 Tuft细胞和ILC2 KO宿主加速了GVHD。我们 假设IL-25效应是由于ILC2的直接刺激或供体IL-17RB+ T引起的 细胞。 意义。 R37扩展阶段的研究将提供有关机制的基本信息 通过哪些对ST2+宿主ILC2和调节性T细胞的影响，通过该peri-BMT IL-33降低了GVHD致死性， 两者都会产生ARE，以及在ISC和Paneth细胞之间发生的EGF/IL-33环路，导致 小肠修复。此外，宿主ILC2在制服GVHD中的关键作用，BMT后ILC2的性质 BMT调节方案和患者易患GVHD之后发生的不足，并且 簇细胞或其产物的基本要求，IL-25将是阐明的。 Lasuy，关键见解将是 关于前LLC2S产生，肠道迁移或分化的无法获得的。 翻译影响：供体和第三方ILC2输注在预防和治疗GVHD方面的效率 支持我们计划的人类ILC2临床试验，该试验将通过其他主持人资助，这些试验将注入第三名 Party iLC2治疗类固醇难治性肠道GVHD，预后预后特别差，在2-中 明尼苏达大学和UNC-CH的机构研究。

项目成果

Molecular modification of a recombinant anti-CD3epsilon-directed immunotoxin by inducing terminal cysteine bridging enhances anti-GVHD efficacy and reduces organ toxicity in a lethal murine model.

• DOI: 10.1182/blood.v96.3.1157
• 发表时间: 2000-08
• 期刊: Blood
• 影响因子: 20.3
• 作者: D. Vallera;D. Kuroki;A. Panoskaltsis‐Mortari;D. Buchsbaum;B. Rogers;B. Blazar

PD-1 restraint of regulatory T cell suppressive activity is critical for immune tolerance.

• DOI: 10.1084/jem.20182232
• 发表时间: 2021-01-04
• 期刊: The Journal of experimental medicine
• 作者: Tan CL;Kuchroo JR;Sage PT;Liang D;Francisco LM;Buck J;Thaker YR;Zhang Q;McArdel SL;Juneja VR;Lee SJ;Lovitch SB;Lian C;Murphy GF;Blazar BR;Vignali DAA;Freeman GJ;Sharpe AH
• 通讯作者: Sharpe AH

Current Concepts and Advances in Graft-Versus-Host Disease Immunology.

• DOI: 10.1146/annurev-immunol-102119-073227
• 发表时间: 2021-04-26
• 期刊: Annual review of immunology
• 影响因子: 29.7
• 作者: Hill GR;Betts BC;Tkachev V;Kean LS;Blazar BR
• 通讯作者: Blazar BR

Amphiregulin in intestinal acute graft-versus-host disease: a possible diagnostic and prognostic aid.

• DOI: 10.1038/s41379-018-0170-z
• 发表时间: 2019-04
• 期刊: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc
• 作者: Amin K;Yaqoob U;Schultz B;Vaughn BP;Khoruts A;Howard JR;DeFor TE;Forster C;Meyer C;Gandhi I;Weisdorf DJ;Rashidi A;MacMillan ML;Blazar BR;Panoskaltsis-Mortari A;Holtan SG
• 通讯作者: Holtan SG

Attenuation of acute graft-versus-host disease in the absence of the transcription factor RORγt.

• DOI: 10.4049/jimmunol.1200858
• 发表时间: 2012-08-15
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Fulton LM;Carlson MJ;Coghill JM;Ott LE;West ML;Panoskaltsis-Mortari A;Littman DR;Blazar BR;Serody JS
• 通讯作者: Serody JS