Project Summary
Neuropathic pain is a major healthcare concern in the US due to its prevalence and high-cost through
healthcare treatment and lost wages. Molecular techniques have been utilized on human dorsal root ganglia
(hDRG) to better understand mechanisms underlying neuropathic pain to lead to the development of novel
therapeutics. This work has demonstrated a sex-specific neuroimmune mechanism in which females show
increased expression of interferon gamma (IFNγ) induced genes. IFNγ is a pro-inflammatory cytokine known
for its anti-viral effects which recently has been shown to produce neuro-inflammation and pain. In animal
models, intrathecal delivery of IFNγ produces paw hypersensitivity, and animals lacking the IFNγ receptor
(IFNGR) fail to produce hyperalgesia following induction of neuropathic pain. Thus, this combined data
suggests a potential female specific role of IFNγ in the hDRG to produce neuropathic pain. Elucidation of IFNγ
mechanisms can lead to the production of sex-specific therapeutics for the treatment of neuropathic pain. I
propose to use hDRG neurons to examine the role of IFNγ in the production of changes consistent with the
development of neuropathic pain. Specific Aim 1 will explore which immune cells present in hDRGs are
capable of producing IFNγ. I will do this through fluorescently activated cell sorting of hDRG neurons of both
sexes to isolate CD4 and CD 8 T cells and monocytes. Then, I will perform RNA sequencing and proteomic
assessments to determine which immune cells possess genes and proteins responsible for IFNγ production
and if any sex differences exist. Then, I will perform a ligand-receptor interactome analysis with the RNA
sequencing and proteomic data sets of each immune cell subtype. This will lead to the generation of testable
hypotheses through which each immune cell can interact with hDRG neurons to lead to the production of
neuropathic pain and if differences exist between the sexes. In Specific Aim 2, I will perform a comprehensive
assessment of the impact of IFNγ signaling on hDRG neurons. In these experiments, I will culture hDRG
neurons of each sex and expose them to IFNγ. First, I will look at the functional impact of IFNγ on hDRG
neurons by using calcium imaging to see if IFNγ can excite and/or sensitize hDRG neurons to later
prostaglandin exposure. Then, I will again expose cultured hDRG neurons to IFNγ and isolate RNA and protein
to perform bulk and single cell RNA sequencing and phosphoproteomic assessment. This will give
unprecedented insight into how hDRG neurons respond functionally, transcriptionally, and proteomically to
IFNγ exposure and if any differences exist between the sexes. In sum, this work can lead to the production of
novel, sex-specific therapeutics for the treatment of neuropathic pain. This work will also provide substantial
scientific and bioinformatic training vital to me reaching my long-term career goal of becoming an independent,
translational researcher as a PI at a large academic institution conducting next generation sequencing and
other omic experiments on human tissue.
