Type 2 cytokines and innate lymphoid cells in pediatric ulcerative colitis

小儿溃疡性结肠炎中的 2 型细胞因子和先天淋巴细胞

• 批准号: 10596871
• 负责人: Michael J Rosen
• 金额: $ 44.6万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-18 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10596871
• 关键词: Address; Adrenal Cortex Hormones; American; Autologous; Biological; Biopsy; Caring; Cell Differentiation process; Cell Therapy; Cells; Child; Childhood; Chronic; Clinical; Coculture Techniques; Colitis; Colon; Crohn' s disease; Custom; Data; Data Set; Diagnosis; Diagnostic; Differentiation and Growth; Disease; Disease remission; Epithelial; Flow Cytometry; Gene Expression; Gene Expression Profile; Goblet Cells; Growth; Homeostasis; Human; IL5 gene; Immune; Immune Response Genes; Immune response; Immunologics; Immunophenotyping; Inflammation; Inflammatory; Interleukin-13; Interleukin-17; Large Intestine; Lymphocyte; Lymphoid Cell; Maintenance; Measures; Mediating; Microfluidics; Modeling; Morbidity - disease rate; Mucins; Mucous Membrane; Mucous body substance; Mus; Outcome; Oxazolone; Pathway interactions; Patients; Pediatric ulcerative colitis; Pharmaceutical Preparations; Production; Prognostic Factor; RNA; Source; Steroids; System; T-Lymphocyte; Testing; Therapeutic; Time; Tissues; Treatment outcome; Ulcerative Colitis; base; clinical predictors; clinical remission; cohort; cytokine; dextran sulfate sodium induced colitis; disorder control; epithelial repair; follow-up; healing; improved; individualized medicine; intestinal epithelium; mesenteric lymph node; microbial colonization; microbiome; mouse model; peripheral blood; predicting response; predictive signature; preservation; prognostic value; prospective; rectal; repaired; response; single-cell RNA sequencing; therapy development; transcriptome; treatment response

PROJECT SUMMARY This proposal addresses two critical gaps in the treatment of pediatric ulcerative colitis (UC). The first gap is the need for patient-specific immunologic profiles that inform precision UC treatments to achieve remission efficiently on the safest medication. The second gap is the need to investigate therapeutic mechanisms that promote epithelial homeostasis and, thereby, improve mucosal healing, one of the best predictors of sustained remission in UC. We previously demonstrated that treatment naïve pediatric UC can be distinguished from colon-only Crohn's disease by increased mucosal expression of type 2 and type 17 immune response genes, as measured by a custom real time RT-qPCR microfluidic array. Furthermore, we observed that high mucosal expression of the type 2 cytokines IL13 and IL5 was associated with 5-fold increased odds of 12-month clinical remission in pediatric UC. We have also shown that IL33, a cytokine that induces type 2 cytokine production by innate lymphoid cells (ILCs), is increased in pediatric UC and is protective in oxazolone colitis in mice, in part through preservation of mucin-producing goblet cells. Maintenance of the mucus layer is critical for epithelial barrier function and mucosal healing. Our preliminary data supports that IL33 induces goblet cell differentiation by promoting the production of IL13 by group 2 ILCs (ILC2s). Our overarching hypothesis is that the induction of mucosal type 2 cytokines by ILC2s in a subset of pediatric UC patients protects the epithelium in the setting of uncontrolled type 17 inflammation and leads to superior treatment outcomes. In Aim 1, we will apply our array to tissues from treatment-naïve patients in a large prospective pediatric UC inception cohort and validate the type 2 gene expression signature for predicting treatment-specific clinical and endoscopic outcome. We will also integrate our array data with microbiome and transcriptome data sets from this cohort to identify key associations between type 2 immunophenotype and microbial colonization/function and host epithelial homeostasis/repair pathways, respectively. In Aim 2, we will identify ILC2s as a key source of type 2 cytokines in pediatric UC using multicolor flow cytometry and single-cell RNA sequencing. We will also determine the effect of autologous peripheral blood ILC2s on human primary colonoid growth and differentiation using a colonoid-immune cell co-culture system. In Aim 3, we will determine the effect of ILC2s on epithelial repair and differentiation, and treatment response during chronic colitis in mice using the T cell transfer and chronic DSS models. Upon completion of Aim 1, we will have validated an assessment of UC immunophenotype for predicting clinical remission and mucosal healing in response to specific treatments in a large well-defined UC inception cohort, which will inform precision patient-specific treatment in UC. Upon completion of Aim 2 and 3, we will have determined whether ILC2s are capable of promoting epithelial growth and goblet cell differentiation in humans and relevant murine models of chronic colitis. This deliverable would support the development of therapies that promote ILC2s or ILC2 cellular therapy for advancing mucosal healing in UC.

项目摘要 该提案解决了小儿溃疡性结肠炎（UC）治疗中的两个关键空白。第一间隙 需要患者特异性免疫学特征，为精确的UC治疗提供信息，以实现缓解 最安全的药物。第二个差距是需要研究治疗机制， 促进上皮内环境稳定，从而改善粘膜愈合，这是持续性炎症的最佳预测因子之一。 UC缓解。我们先前证明，未接受过治疗的儿童UC可与 通过增加2型和17型免疫应答基因的粘膜表达， 如通过定制的真实的时间RT-qPCR微流体阵列测量的。此外，我们观察到高粘膜 2型细胞因子IL 13和IL 5的表达与12个月临床复发率增加5倍相关。 儿童UC的缓解。我们还表明，IL 33，一种通过免疫调节诱导2型细胞因子产生的细胞因子， 先天性淋巴样细胞（ILC）在儿童UC中增加，并在小鼠恶唑酮结肠炎中具有保护作用， 通过保存产生粘蛋白的杯状细胞。粘液层的维持对于上皮细胞的生长至关重要。 屏障功能和粘膜愈合。我们的初步数据支持IL 33诱导杯状细胞分化 通过促进第2组ILC（ILC 2）产生IL 13。我们的首要假设是， 在儿童UC患者亚组中，ILC 2的粘膜2型细胞因子在环境中保护上皮 不受控制的17型炎症，并导致上级治疗结果。在目标1中，我们将应用我们的 在一个大型前瞻性儿童UC初始队列中，将阵列与来自初治患者的组织进行比较，并验证 用于预测治疗特异性临床和内镜结果的2型基因表达特征。我们将 我还将我们的阵列数据与来自该队列的微生物组和转录组数据集相结合， 2型免疫表型与微生物定植/功能和宿主上皮细胞之间的关系 稳态/修复途径。在目标2中，我们将确定ILC 2作为2型细胞因子的关键来源 在儿科UC中使用流式细胞术和单细胞RNA测序。我们还将确定 自体外周血ILC 2对人原代结肠样细胞生长和分化的影响 类集落-免疫细胞共培养系统。在目标3中，我们将确定ILC 2对上皮修复的影响， 在小鼠中使用T细胞转移和慢性DSS在慢性结肠炎期间的治疗反应 模型完成目标1后，我们将验证UC免疫表型的评估， 预测明确的大型UC患者对特定治疗的临床缓解和粘膜愈合 初始队列，这将为UC的精确患者特异性治疗提供信息。在完成目标2和3后， 我们将确定ILC 2是否能够促进上皮细胞生长和杯状细胞增殖。 在人类和慢性结肠炎的相关鼠模型中的分化。该交付品将支持 开发促进ILC 2或ILC 2细胞疗法的疗法，以促进UC的粘膜愈合。

项目成果

Sprouty2 limits intestinal tuft and goblet cell numbers through GSK3β-mediated restriction of epithelial IL-33.

• DOI: 10.1038/s41467-021-21113-7
• 发表时间: 2021-02-05
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Schumacher MA;Hsieh JJ;Liu CY;Appel KL;Waddell A;Almohazey D;Katada K;Bernard JK;Bucar EB;Gadeock S;Maselli KM;Washington MK;Grikscheit TC;Warburton D;Rosen MJ;Frey MR
• 通讯作者: Frey MR