Single cell analytics and drug discovery at the endometrial-myometrial junction in symptomatic adenomyosis - Adenomyosis is a common, understudied, estrogen-dependent uterine disorder that greatly impacts the well-
being of women. It is characterized by endometrial cells in the myometrium that elicit inflammation and smooth
muscle hyperplasia and hypertrophy, and symptoms of severe dysmenorrhea and heavy menstrual bleeding
(HMB), subfertility, and poor pregnancy outcomes. The disease is believed to derive from physiologic
(pregnancy) or iatrogenic (uterine surgery, endometrial curettage) trauma to the endometrial-myometrial
junction (EMJ), with local inflammation promoting migration of endometrial basalis into the myometrium.
Therapies include suppressing estrogen production and action, but long-term efficacy and acceptability of these
are limited, and disease excision is complicated, and while hysterectomy is curative, it is unacceptable in
patients trying to conceive. The goal of this proposal is to gain insights into adenomyosis pathophysiology at
single cell resolution, leading to novel and targeted medical therapies to abate symptoms and improve quality
of life of those affected. Our recent bulk RNAseq of endometrial and sub-endometrial inner myometrial
specimens from patients with diffuse adenomyosis vs controls support collective endometrial cell migration and
reveal marked endometrial inflammation, and myometrial neuroactive ligand-receptor and cytokine-receptor
interactions. In preliminary studies, we developed and applied a bioinformatic drug discovery pipeline to these
data and identified novel and known compounds of interest (COI) that reverse disease networks in silico. We
hypothesize that unique features in the EMJ give rise to adenomyosis and its symptoms, and that EMJ
homeostasis can be restored using novel or re-purposed drugs discovered by expression reversal of disease
networks. In Aim 1, we will create an EMJ compartment-specific single cell transcriptomic atlas from 40 well-
annotated samples from uteri with and without diffuse adenomyosis across the menstrual cycle, archived in our
Human Uterine Research Biorepository and with ongoing prospective accrual as needed. In Aim 2, we will
apply a transcriptomics based drug repurposing pipeline to publicly available and newly generated uterine
transcriptome datasets to identify COI to reverse the adenomyosis phenotype. Our multidisciplinary,
collaborative team has vast expertise in uterine disorders, omics technologies, molecular and cell biology,
immunology, and computational sciences. The impact of our proposal, using advanced technologies and high
order analytics, is to elucidate cellular dysfunction of EMJ endometrium and inner myometrium at single cell
resolution in adenomyosis patients and discover drugs for specific cell type targets for future studies for disease
regression and symptom abatement.
