Functional consequences of putative analgesic receptor activation in human DRG neuronal cultures from healthy donors

National Center For Advancing Translational Sciences

Investigators

Abstract

One of the highest priorities for the pain field is the development of novel non-opioid orally bioavailable pain medications. The ongoing opioid crisis demonstrates the need for alternatives to the opioid class of drugs. Attempts to avoid opioids because of their deleterious peripheral and central side effects, including potentially lethal respiratory depression, and the stigma associated with opioid use has led to a high rate of pain undertreatment. One strategy to avoid central side effects such as addiction is to inactivate the sensory afferent ending by targeting dorsal root ganglion neurons. Clinical investigations and spatial transcriptomic characterization by the Mannes lab of the DRG have recently uncovered all of the targetable receptors expressed in the most clinically relevant population of DRG neurons. These neurons co-express transient receptor potential vanilloid 1 (TRPV1) and the mu-opioid receptor (OPRM1). The current study aims to interrogate a select number of targets identified in human pain models and transcriptomics that are hypothesized to be highly likely to modulate the TRPV1+/OPRM1+ neurons. These studies will form the basis to demonstrate analgesic actions of these novel targets in a simplified human neuronal culture model, that will provide support for future studies in vivo. Ultimately, these investigations aim to identify novel non-opioid analgesics suitable for Phase I safety trials in human and veterinary pain indications. The primary focus of SCTL scientists in the past year has been to utilize rigorous and efficient iPSC differentiation protocols for a variety of cell types relevant for both disease modeling and cell therapy applications, hypothalamic arcuate neurons (e.g., T2D), nociceptors (e.g., pain research), cortical neurons (e.g., Tay Sachs disease), astrocytes (e.g., free sialic acid storage disease), insulin-producing cells (e.g., type 1 diabetes), hepatocytes (e.g., liver failure), trophectoderm (e.g., placental development), cerebellar organoids (e.g., Friedreich’s ataxia), and dorsal root ganglion organoids (e.g., chemotherapy induced peripheral neuropathy). In FY25, our lab published six manuscripts. One manuscript demonstrates the effects of satellite glia on iPSC-derived sensory neuron differentiation and maturation. The second manuscript demonstrates the translational application of iPSC-derived hypothalamic arcuate neurons for the identification of environmental compounds that may trigger early female puberty. The third manuscript demonstrates a highly efficient protocol for the generation of iPSC-derived trophoblast. The fourth manuscript establishing a scalable differentiation platform for the generation of hypothalamic arcuate neurons from iPSCs. The fifth manuscript is a review article in collaboration with the international stakeholders on the promotion of best practices for stem cell research. The sixth manuscript establishes a protocol for the generation of cerebellar organoids and their application in Friedreich’s ataxia disease modeling. In this collaboration, the SCTL mined our bulk RNA-Seq data to demonstrate that specific GPR targets are expressed in our hPSC-derived sensory neurons, these cells were then generated for validation studies in the Mannes lab. Ongoing Collaborations: 1) Shuibing Chen (Cornell University): Multi-omic characterization of type 2 diabetes patient iPSC-derived cell types collaboration between NCATS, NHGRI, Cornell and Columbia 2) Claudia Doege (Columbia University): Multi-omic characterization of type 2 diabetes patient iPSC-derived cell types collaboration between NCATS, NHGRI, Cornell and Columbia 3) Lauretta Lacko (Cornell University): Multi-omic characterization of type 2 diabetes patient iPSC-derived cell types collaboration between NCATS, NHGRI, Cornell and Columbia 4) Robert Schwartz (Cornell University): Multi-omic characterization of type 2 diabetes patient iPSC-derived cell types collaboration between NCATS, NHGRI, Cornell and Columbia 5) Ronald Khan (Harvard University): Multi-omic characterization of type 2 diabetes patient iPSC-derived cell types collaboration between NCATS, NHGRI, Cornell and Columbia 6) Stephan Parker (University of Michigan): Multi-omic characterization of type 2 diabetes patient iPSC-derived cell types collaboration between NCATS, NHGRI, Cornell and Columbia 7) Fei Wang (Cornell University): Multi-omic characterization of type 2 diabetes patient iPSC-derived cell types collaboration between NCATS, NHGRI, Cornell and Columbia 8) Leslie Thompson (University of California, Irvine): Huntington’s disease modeling using functional genomics and hPSC-derived astrocytes 9) Daniel Paull (New York Stem Cell Foundation): Role of TCF7L2 in beta cell differentiation and function using iPSCs collaboration between NCATS, NHGRI and NYSCF. 10) Thomas Kent (Texas A&M University): Pleiotropic oxidized carbon nanozymes as new therapeutics for Friedreich’s ataxia treatment 11) Sang Jin Lee (Wake Forest Institute of Regenerative Medicine): Generation of human 3D skeletal muscle composite to study muscle pathophysiology 12) David Bennett (University of Oxford): Transcriptomic analysis of diabetic peripheral neuropathy patient iPSC-derived cell types 13) Anna Moreno (Navega Therapeutics): Testing a Reversible Gene Editing Method for Analgesia using iPSC-derived Sensory Neurons 14) Rosalind Segal (Harvard University): Studies of Chemotherapy Induced Peripheral Neuropathy (CIPN) in iPSC derived peripheral sensory neurons 15) Martin Schneider (University of Maryland Baltimore): Electrophysiological characterization of iPSC-derived neural cell types 16) Clifford Woolf (Harvard): Characterization of iPSC-derived nociceptors Collaboration between SCTL and Harvard 17) Bruce Bean (Harvard University): Characterization of iPSC-derived nociceptors Collaboration between SCTL and Harvard 18) Erick Hernandez-Ochoa (University of Maryland Baltimore): Electrophysiological characterization of iPSC-derived neural cell types 19) Jun-Ho La (University of Texas Medical Branch): Role of GPR37 in pain memory erasure in iPSC-derived sensory neurons. 20) Laura Pollard (Greenwood Genetic Center): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 21) Richard Steet (Greenwood Genetic Center): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 22) Raymond Wang (Children’s Hospital of Orange County): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 23) Monkol Lek (Yale University): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 24) Kostantin Dobrenis (Albert Einstein College of Medicine): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 25) Steven Walkley (Albert Einstein College of Medicine): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 26) Christine Anne-Longin (University of Paris): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 27) Bruno Gasnier (University of Paris): Preclinical development of potential gene editing therapy for Free Sialic Acid Storage Disorder collaboration between NCATS and the FSASD Consortium 28) Ross Marklein (Federal Drug Administration): Development of a platform for manufacturing iPSC-derived mesenchymal stromal cells producing extracellular vesicles for neurodegenerative disease