Elucidating the role of Prdm12 in the development and maintenance of nociceptive neurons

阐明 Prdm12 在伤害性神经元发育和维持中的作用

基本信息

批准号：
10019343
负责人：
Mark Landy
金额：
$ 3.18万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-12 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10019343
关键词：
AVIL gene Accounting Acute Pain Adult Afferent Neurons Analgesics Area Bacterial Artificial Chromosomes Behavioral Biological Assay Caliber Cell Death Chromatin Code Congenital Pain Insensitivity DNA Sequence Alteration Defect Development Disease model Economic Burden Electrophysiology (science)Embryo Embryonic Development Enterobacteria phage P1 Cre recombinase Epidemic Esthesia Gene Expression Gene Expression Profile Genes Genetic Transcription Goals Human Impairment Knock-out Knockout Mice Lead Maintenance Mechanics Medical Medicine Mitotic Modality Modeling Molecular Morbidity - disease rate Motor Mus Mutant Strains Mice Mutation NTRK1 gene Neurons Nociception Nociceptive Stimulus Nociceptors Opioid Overdose Pain Pain Research Painless Pathogenesis Pathway interactions Patients Phenotype Play Population Prevalence Productivity Proprioception Proteins Pruritus Research Role Sensory Series Sodium Channel Spinal Ganglia Tamoxifen Testing Touch sensation Transgenes Variant behavioral phenotyping behavioral study cell type chronic neuropathic pain chronic painful condition clinical pain conditional knockout cost drug discovery economic cost insight loss of function loss of function mutation mRNA sequencing mortality mouse model neurogenesis neurotrophic factor non-opioid analgesic novel novel strategies pain behavior pain sensation postnatal transcription factor vibration voltage

项目摘要

PROJECT SUMMARY Painful conditions are a huge medical burden with an estimated prevalence of 32% per year and an economic burden over $600 billion accounting for treatment and lost productivity costs. While opioids can provide analgesic relief for some patients, their increasing use in medicine led to an epidemic of its own, with an ever-increasing mortality rate due to overdose, highlighting the need for novel, non-opioid analgesics. Recently, research to meet this need has been fueled by the study of mutations leading to painlessness (Congenital Insensitivity to Pain (CIP)). As part of this effort, I have focused on a newly identified variant of CIP caused by a series of loss-of- function mutations in the chromatin-modifying factor, PRDM12. Patients with Prdm12 mutations are unable to detect painful (nociceptive) stimuli, without defects in other sensory modalities (touch, vibration, proprioception, and itch). Furthermore, Prdm12 has been proposed to play a role in nociceptor development, consistent with the finding that humans with PRDM12-associated CIP lack small diameter nociceptors responsible for transmitting pain sensation. To better understand how Prdm12 loss-of-function leads to CIP, I developed a mouse model of Prdm12-associated CIP to test the hypothesis that PRDM12 expression is required for the development and maintenance of nociceptive sensory neurons. In Aim 1, I will use this model to analyze the developmental and behavioral defects that result from Prdm12 loss-of-function mice compared to controls. I will start by looking for differences in neurogenesis, cell type specification, or cell death in dorsal root ganglia of Prdm12 loss-of-function mice. I will then compare these developmental changes with behavioral studies investigating nociceptive sensation of these mice to understand how loss of particular subsets of sensory neurons leads to defects in specific modalities of nociception. Finally, I will use mRNA-seq to begin to investigate the mechanism of action of PRDM12 at various stages of development. In Aim 2, I will test whether PRDM12 is involved in the maintenance of nociceptive sensory neurons. I will do this by using a tamoxifen-inducible model to knockout Prdm12 at late embryonic to adulthood stages, and examining the DRG composition, pain behavior phenotype, and transcriptional changes in these mice. Additionally, I will use cultured DRG neurons from these mice to assess changes in neuronal populations and excitability using electrophysiology. Overall, the project will provide new insight into the developmental role of Prdm12 and our understanding of pain sensation, while also serving to open potentially new areas of analgesic discovery that could benefit both acute and chronic pain conditions.

项目概要疼痛状况是一项巨大的医疗负担，估计每年患病率达 32%，并且会造成经济损失治疗和生产力损失的成本超过 6000 亿美元。虽然阿片类药物可以提供镇痛作用尽管这些药物对一些患者来说是一种缓解，但它们在医学上的使用越来越多，导致了其自身的流行病，并且感染人数不断增加过量服用导致的死亡率，凸显了对新型非阿片类镇痛药的需求。最近研究会对导致无痛的突变的研究推动了这种需求（先天性对疼痛不敏感）（CIP））。作为这项工作的一部分，我重点研究了一种新发现的 CIP 变体，该变体是由一系列丢失- 染色质修饰因子 PRDM12 的功能突变。患有 Prdm12 突变的患者无法检测疼痛（伤害性）刺激，而其他感觉方式（触摸、振动、本体感觉、和痒）。此外，Prdm12 已被提议在伤害感受器发育中发挥作用，这与发现患有 PRDM12 相关 CIP 的人类缺乏负责传播的小直径伤害感受器疼痛感。为了更好地了解 Prdm12 功能丧失如何导致 CIP，我开发了一个小鼠模型 Prdm12 相关的 CIP 来检验 PRDM12 表达对于伤害性感觉神经元的发育和维持。在目标1中，我将使用这个模型来分析与对照组相比，Prdm12 功能丧失小鼠导致的发育和行为缺陷。我首先寻找神经发生、细胞类型规范或背根神经节细胞死亡的差异 Prdm12 功能丧失小鼠。然后我会将这些发育变化与行为研究进行比较研究这些小鼠的伤害性感觉，以了解特定感觉子集的丧失是如何发生的神经元导致特定伤害感受方式的缺陷。最后，我将使用mRNA-seq开始研究 PRDM12在不同发育阶段的作用机制。在目标 2 中，我将测试 PRDM12 是否是参与伤害性感觉神经元的维持。我将通过使用他莫昔芬诱导模型来做到这一点在胚胎晚期到成年阶段敲除 Prdm12，并检查 DRG 组成、疼痛行为这些小鼠的表型和转录变化。此外，我将使用来自这些的培养 DRG 神经元使用电生理学评估小鼠神经元群体和兴奋性的变化。总体而言，该项目将为 Prdm12 的发育作用和我们对疼痛感觉的理解提供了新的见解，同时也有助于开辟镇痛药物发现的潜在新领域，使急性和慢性疼痛受益状况。