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High content analgesic screening from human nociceptors

从人类伤害感受器中筛选高含量镇痛剂

基本信息

批准号：
10578042
负责人：
GREGORY O DUSSOR
金额：
$ 37.02万
依托单位：
UNIVERSITY OF TEXAS DALLAS
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：
2023-02-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10578042
关键词：
Absence of pain sensation Action Potentials Acute Pain Address Afferent Neurons Agonist Analgesics Anatomy Animals Baclofen Basic Science Behavior Biochemical Biological Assay Biology Capsaicin Cell physiology Cells Chili Pepper Data Development Electrodes Electrophysiology (science)Ethics Evaluation Event Failure Genes Heterogeneity Home Homo sapiens Human In Vitro Inflammatory Interleukin 6 Receptor Interleukin-6 Libraries Measures Methods Modeling Modernization Molecular Target Mus National Cancer Institute Natural Products Natural Source Neurons Nociception Nociceptors Operative Surgical Procedures Organ Donor Organism Pain Paper Patients Peripheral Pharmacology Phase Phenotype Physiological Physiology Plants Process Assessment Proteins Publishing Rattus Research Rodent Rodent Model Sorting Source Speed Spinal Ganglia Stimulus TRPV1 gene Technology Testing Therapeutic Translating Translations antagonist chronic pain cytokine efficacy study falls high throughput screening in vivo induced pluripotent stem cell inflammatory pain inhibitor innovation marine organism mouse model multi-electrode arrays novel opioid epidemic pain model patch clamp pharmacologic pharmacophore pre-clinical repository screening single cell sequencing success tool transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY The opioid crisis has driven home the need for new, non-addictive analgesics to treat both severe acute and chronic pain. Over the past 25 years, the search for such novel analgesics has had only a few successes and many failures. There are two drawbacks of modern analgesic screening that we aim to correct with our screening assay. First, screening for novel analgesics typically relies on a biochemical readout that is readily amenable to high throughput screening (HTS). While such an approach has advantages (e.g., speed), it does not account for the complexity and heterogeneity of nociceptor sensory neuron biology. Second, many new analgesic targets and efficacy studies from HTS rely on rodent behavior and cellular physiology. For clear ethical reasons, rodent efficacy studies have made complete sense, and yet, the differences between homo sapiens and murine organisms have led to a failure to translate exciting preclinical targets to human pain. The mechanisms for these failures are now being illuminated as multiple research groups, including our own, have begun to study sensory neuron physiology and pharmacology in primary human dorsal root ganglia (DRG). Differences in expression of “standard” nociceptor markers between mice, rats, and humans illustrate the importance of moving into human cells as early as possible in the development of novel analgesics. In this proposal, we address both issues above by performing screening of natural product fractions for their ability to reduce hyperexcitability in human induced pluripotent stem cell-derived nociceptors. In the past, the promise of derived nociceptors has fallen short of true human nociceptor replication. Based on our preliminary and published data, we believe that our cells, RealDRGs, truly are “nociceptors.” In this proposal, we will combine this abundant, replicable, and reliable source of human “nociceptors” with cutting-edge multi-electrode array (MEA) technology to screen natural product fractions for analgesic-like effects. Over the last several years, we have used mouse DRGs with MEAs to show proof-of- concept (PoC) analgesic screening including documenting and validating a Z’-factor assay quality assessment process. We will use the R61 portion to determine (1) the optimal stable baseline after plating of RealDRGs to estimate the active electrode yield, (2) the stability in firing rate after exogenous sensitization, (3) the Z’ factor, and (4) PoC screening of known nociceptor inhibitors. Upon reaching our quantitative milestones, we will then progress in the R33 phase to (1) screen natural product fractions from the National Cancer Institute with (2) final testing of subfractions in RealDRGs and in human primary DRGs.

项目总结