Schwann Cell-derived neuro-gliogenesis

雪旺细胞衍生的神经胶质细胞生成

• 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
• 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

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.

总结 肠神经系统（ENS）是一个复杂的神经嵴源性神经元和神经胶质细胞网络， 用于调节关键的肠道功能，包括运动、感觉和分泌。不幸的是，ENS 经常遭受导致运动和其他异常的损伤。通常，这会导致衰弱性疾病 几乎没有可用的治疗方案。令人兴奋的是，现在有越来越多的证据表明，出生后ENS损伤引起的 神经发生重要的是，通过对成年动物模型的研究，我们已经证明许旺细胞（SC）可以 与外源性神经一起进入肠道，然后分化为特定的神经元和神经胶质亚型 （肠神经胶质生成）。因此，SC提供了一种意想不到的细胞来源，以重新填充受损的神经元， 肠神经胶质。此外，我们发现微生物组操纵是诱导雪旺氏病的有力方法。 细胞介导的肠神经胶质细胞生成导致ENS的功能恢复，这是通过 血清素5 HT 4通路。然而，出生后ENS神经胶质发生的许多方面并不完全 了解，包括从SC神经胶质发生的功能影响，和治疗潜力， 用于人类疾病中的5 HT 4操纵，旨在增强SC诱导的神经胶质再生。 基于我们发表的小鼠和人类的初步结果，我们的总体假设 SC从肠道的外部神经支配迁移到肠道是出生后 肠神经胶质细胞生成，并且这种ENS再生反应由微生物组通过以下途径调节： 5 HT4。为了验证这一新的假设，我们提出：目的1将表征出生后SC衍生的肠神经元， 使用可诱导的荧光标记的小鼠在微生物组根除/重建后的胶质细胞生成。我们将 还通过广泛的体内运动性测定确定SC神经胶质发生的功能作用， 渗透性和使用钙成像的细胞功能的离体表征。此外，我们将 使用白喉毒素小鼠模型确定消除进入肠道的SC的功能效果。在 目的2，我们将使用两个敲除小鼠系：（1）P0 CreER/tdT：：Tph 1-/-和（2）P0 CreER/tdT：：Tph 2-/-， 确定血清素的来源和我们的研究结果可能的临床应用，通过评估SC 对5 HT 4激动剂普卢卡必利的反应。我们还将确定特定的代谢组学和转录组学概况 胃肠道（粘膜和肌间隔室）。最后，在目标3中，我们将确定人体的成分 微生物群-宿主串扰调节慢结肠患者SC源性肠神经胶质细胞生成 运输/运动障碍包括5 HT 4激动剂的作用（即，普卢卡必利、替加色罗）对ENS完整性/神经元的影响， 胶质细胞再生和功能，并确定宏基因组概况在我们的患者队列。最后，我们将表演 将来自这些受试者的粪便移植到无菌（GF）小鼠中以评估ENS恢复。结果从这个 该提案将是持续推进靶向再生治疗的关键， 先天性和后天性神经肠道疾病。