Schwann Cell-derived neuro-gliogenesis

雪旺细胞衍生的神经胶质细胞生成

基本信息

批准号：
10735664
负责人：
Jaime Belkind-gerson
金额：
$ 48.86万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10735664
关键词：
Ablation Address Affect Age Agonist Anatomy Animal Model Antibiotics Bacteria Biological Assay Biopsy Bromodeoxyuridine Calcium Cell Death Cell Proliferation Cell physiology Cells Cessation of life Clinical Coculture Techniques Colon Complement Complex Diagnosis Diphtheria Toxin Disease Dryness Enteral Enteric Nervous System Esthesia Excision Feces Gastrointestinal Diseases Gastrointestinal Injury Gastrointestinal Motility Gastrointestinal tract structure Germ-Free Glial Differentiation Health Human Human Microbiome Image In Situ Nick-End Labeling In Vitro Inflammation Injury Intervention Intestinal Diseases Intestinal permeability Intestines Knockout Mice Knowledge Kynurenine Label Life Location Malignant Neoplasms Maps Measurement Measures Mediating Metagenomics Methods Migrating Myoelectric Complex Mission Modeling Motor Mucous Membrane Mus Myelin P0 Protein Natural regeneration Nerve Nerve Regeneration Nervous System Trauma Neural Crest Neuroglia Neuronal Injury Neurons Output Pathway interactions Patients Permeability Prevention Process Proliferating Public Health Publishing Quality of life Recovery Recovery of Function Regenerative response Reporter Research Role Schwann Cells Sensory Disorders Serotonin Signal Transduction Signaling Molecule Solid Source Streptococcus sanguis System Testing Therapeutic Thick Tryptamines United States National Institutes of Health Work Zebrafish aged biomarker identification cell motility clinical application cohort conditional knockout density disability experimental study fecal transplantation gastrointestinal gastrointestinal function gliogenesis gut microbiota host microbiome human disease improved in vivo injury recovery innovation mature animal metabolomics microbiome microbiota motility disorder mouse model nerve stem cell nerve supply neurogenesis novel novel therapeutic intervention postnatal pre-clinical prenatal promoter receptor regenerative therapy response sex tegaserod transcriptomics translational study

项目摘要

SUMMARY The enteric nervous system (ENS) is a complex network of neural crest-derived neurons and glia responsible for regulating key intestinal functions including motility, sensation, and secretion. Unfortunately, the ENS is frequently subject to injury leading to motor and other abnormalities. Often, this leads to debilitating disorders with few available treatment options. Excitingly, there is now mounting evidence of postnatal ENS injury-induced neurogenesis. Importantly, through work on adult animal models we have shown that Schwann cells (SC) can enter the gut alongside the extrinsic nerves and then differentiate into specific neuronal and glial subtypes (enteric neuro-gliogenesis). Thus, SC provide an unexpected source of cells to repopulate injured neurons and enteric glia. Furthermore, we have found that microbiome manipulation is a powerful method to induce Schwann cell-mediated enteric neuro-gliogenesis leading to functional recovery of the ENS and that this is mediated via the serotonin 5HT4 pathway. However, many aspects of postnatal ENS neuro-gliogenesis are not fully understood, including the functional impact of the neuro-gliogenesis from the SC, and the therapeutic potential for 5HT4 manipulation in human disease aiming for an enhanced SC-induced neuro-glial regeneration. Building on our published and preliminary results from mice and humans, our overarching hypothesis is that SC migrating into the gut from the gut’s extrinsic innervation are an important source for postnatal enteric neuro-gliogenesis, and that this ENS regenerative response is regulated by the microbiome via 5HT4. To test this novel hypothesis, we propose: Aim 1 will characterize postnatal SC-derived enteric neuro- gliogenesis after microbiome eradication/re-establishment using inducible, fluorescently labeled mice. We will also determine the functional effects of SC neuro-gliogenesis through extensive in vivo assays of motility and permeability and ex vivo characterization of cellular function using calcium imaging. Additionally, we will determine the functional effect of eliminating the SC entering the gut using a diphtheria toxin mouse model. In Aim 2, we will use two knockout mouse lines: (1) P0CreER/tdT::Tph1-/- and (2) P0CreER/tdT::Tph2-/- to determine the source of serotonin and the possible clinical applications of our findings by evaluating the SC response to a 5HT4 agonist, prucalopride. We will also identify specific metabolomic and transcriptomic profiles of the GI tract (mucosal and myenteric compartments). Finally in Aim 3, We will determine components of human microbiome-host crosstalk regulating SC-derived enteric neuro-gliogenesis in patients with slow colonic transit/dysmotility including the effect of 5HT4 agonists (i.e., prucalopride, tegaserod) on the ENS integrity/neuro- glial regeneration and function and determine metagenomic profiles in our patient cohort. Last, we will perform fecal transplants from these subjects into germ-free (GF) mice to evaluate ENS recovery. Results from this proposal will be key for the continued progress in targeted regenerative therapy for the treatment of congenital and acquired neuro-intestinal disease.

概括肠神经系统 (ENS) 是一个由神经嵴衍生神经元和神经胶质细胞组成的复杂网络，负责不幸的是，ENS 是调节关键的肠道功能，包括蠕动、感觉和分泌。经常受到伤害，导致运动和其他异常，这通常会导致衰弱性疾病。令人兴奋的是，目前有越来越多的证据表明产后 ENS 损伤所致。重要的是，通过对成年动物模型的研究，我们已经证明雪旺细胞（SC）可以与外在神经一起进入肠道，然后分化为特定的神经和神经胶质亚型（肠神经胶质细胞生成）因此，SC 提供了意想不到的细胞来源来重新填充受损的神经元和此外，我们发现微生物组操作是诱导雪旺氏细胞的有效方法。细胞介导的肠神经胶质细胞生成导致 ENS 功能恢复，这是通过然而，出生后 ENS 神经胶质细胞生成的许多方面尚未完全实现。了解，包括 SC 神经胶质生成的功能影响以及治疗潜力用于人类疾病中的 5HT4 操作，旨在增强 SC 诱导的神经胶质细胞再生。基于我们已发表的小鼠和人类的初步结果，我们的总体假设 SC 从肠道的外在神经支配迁移到肠道是产后的重要来源肠道神经胶质细胞生成，并且这种 ENS 再生反应是由微生物组通过调节 5HT4。为了检验这一新假设，我们提出：目标 1 将表征出生后 SC 衍生的肠神经- 使用可诱导的荧光标记小鼠消灭/重建微生物组后的胶质细胞生成。还通过广泛的体内运动测定来确定 SC 神经胶质细胞生成的功能效应此外，我们将使用钙成像进行细胞功能的渗透性和离体表征。使用白喉毒素小鼠模型确定消除 SC 进入肠道的功能效果。目标 2，我们将使用两个基因敲除小鼠品系：(1) P0CreER/tdT::Tph1-/- 和 (2) P0CreER/tdT::Tph2-/- 通过评估 SC 来确定血清素的来源以及我们的发现的可能的临床应用对 5HT4 激动剂普卡必利的反应我们还将确定特定的代谢组学和转录组学特征。最后，在目标 3 中，我们将确定人体的成分。微生物组-宿主串扰调节慢结肠患者 SC 源性肠神经胶质细胞生成转运/运动障碍，包括 5HT4 激动剂（即普卡必利、替加色罗）对 ENS 完整性/神经元的影响最后，我们将进行神经胶质再生和功能并确定患者队列中的宏基因组图谱。将这些受试者的粪便移植到无菌 (GF) 小鼠中以评估 ENS 恢复结果。该提案将是靶向再生疗法持续取得进展的关键先天性和获得性神经肠道疾病。