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.

摘要 我们的总体目标是通过先天性淋巴结转移预防和治疗移植物抗宿主病（GVHD）。 2型细胞（ILC 2）抗炎和组织修复特性。我们发现，宿主ILC 2在 通过全身照射{TBI}或化疗，并保持耗尽>/=90天。这一发现高度 相关，因为外周血活化的ILC 2和GVHD之间存在负相关。就像这样。我们 试图确定是否可以使用成熟供体ILC 2的补充输注来预防鼠 GVHD。我们首次证明了供体ILC 2可以预防或部分治疗双调蛋白中的GVHD。 （AREG）依赖过程。AREG或ILC 2与肠干细胞（ISC）的直接接触是否支持小的 TBI处理的小鼠或类器官中的肠上皮细胞修复是未知的。此外，第三方ILC 2输注 也显著降低鼠GVHD致死率。为了翻译的目的，我们发现ILC 2是 对类固醇有抵抗力BMT（骨髓移植）周IL-33在BMT日增加ST 2/IL 33 R + ILC 2 0和减少GVHD。Ko小鼠具有加速的GVHD;给予预BMT的IL-33防止了完全的ILC 2损失。IL-33 ko受体的上皮细胞增殖低下，ISC和潘氏细胞减少，隐窝高度降低 和数字离体肠类器官培养模型显示，IL-33协调再生， 诱导表皮生长因子（EGF），TBI显著降低。EGF修复了ISC缺陷， ISCs和Paneth细胞之间的肠修复IL-33/EGF环。供体IL-13 ko ILC 2或宿主IL-13 Ra ko小鼠具有 减少GVHD。ILC 2 IL 13支持ST 2+簇状细胞和杯状细胞。簇细胞产生IL-25驱动ILC 2 生产与生存。TBI显著减少了以下的毛簇细胞：38天和ko接收者有显着增加 在GVHD。当给予野生型小鼠外源性IL-25时，显著降低GVHD。科的后果 簇细胞（和ILC 2）对供体T细胞扩增、运输和功能的影响是未知的。IL-25的作用， IL 17 RB未被检测。我们将讨论ILC 2，IL-33和IL-10之间的动态和相互作用。 TBI后和GVHD期间的宿主肠细胞（簇状细胞、ISC、潘氏细胞）。 目的1将检验以下假设：由于ILC 2 BM生态位的破坏，供体ILC 2再增殖失败 支持ILC 2。体外前-ILC 2分化、成熟和扩增±促炎 将研究细胞因子和ILC 2支持细胞因子。GATA 3-GFPhi前ILC 2/成熟ILC 2转移 致死性照射的同基因BMT受体将提供关于重新填充 BM。如果BM不能支持pre-ILC 2/ILC 2，我们将研究干细胞缺陷小鼠。目标2将测试 Pre-ILC 2/ILC 2及其分泌产物对肠细胞亚群具有直接作用。 来自野生型和具有同基因Tcl 3和ILC 2的IL-33 ko小鼠的肠类器官培养物将测量 类器官的大小、数量和与增殖、细胞周期调节和特异性 上皮谱系标志物。抗AREG mAb或与AREG ko共培养 淋巴细胞的特征在于促进上皮再生。目标3将检验以下假设： 细胞通过ILC 2释放IL-13对ILC 2的发育和存活至关重要，IL-13刺激簇细胞， 释放导致ILC 2增殖的IL-25（目的3）。簇状细胞和ILC 2 ko宿主具有加速的GVHD。我们 假设IL-25作用是由于ILC 2的直接刺激或供体IL-17 RB + T的刺激 细胞 意义R37扩展阶段的研究将提供有关机制的基本信息 通过该方法，外周BMT IL-33通过对ST 2+宿主ILC 2和调节性T细胞的作用降低GVHD致死率， 两者都产生AREG，并且在ISCs和Paneth细胞之间发生的EGF/IL-33环导致 小肠修复此外，宿主ILC 2在抑制GVHD中的关键作用，BMT后ILC 2的性质， 在预骨髓移植预处理方案后发生的缺陷，使患者易患GVHD， 将阐明毛丛细胞或其产物IL-25基本要求。Lasuy，批判性的见解将是 获得了关于不能产生pre-ILC 2、肠道迁移或分化的结论。 转化影响：供体和第三方ILC 2输注在预防和治疗GVHD中的疗效 支持我们计划的人类ILC 2临床试验，通过其他赞助，这将注入“现成的”第三 在一项2- 在明尼苏达大学和UNC-CH进行机构研究。

项目成果

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

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

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

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

Chronic GvHD NIH Consensus Project Biology Task Force: evolving path to personalized treatment of chronic GvHD.

• DOI: 10.1182/bloodadvances.2022007611
• 发表时间: 2023-09-12
• 期刊: BLOOD ADVANCES
• 影响因子: 7.5
• 作者: Buxbaum, Nataliya P.;Socie, Gerard;Hill, Geoffrey R.;Macdonald, Kelli P. A.;Tkachev, Victor;Teshima, Takanori;Lee, Stephanie J.;Ritz, Jerome;Sarantopoulos, Stefanie;Luznik, Leo;Zeng, Defu;Paczesny, Sophie;Martin, Paul J.;Pavletic, Steven Z.;Schultz, Kirk R.;Blazar, Bruce R.
• 通讯作者: Blazar, Bruce R.