Abstract Details

Title Pathogenic Th17 Cells in Autoimmune Myasthenia Gravis Exhibit a Pro-inflammatory Transcriptomic Signature that Promotes Disease Pathogenesis

Topic Autoimmune Neurology

Presentation(s) S41 - Autoimmune Neurology 3: Autoimmune Encephalitis, Paraneoplastic Neurologic Disorders, Antibody-Mediated Diseases, Immunology (4:54 PM-5:06 PM)

Poster/Presentation
Number 008

Background A subset of Th17 cells co-produce IFN-γ and IL-17 (pathogenic Th17 cells) that are implicated as a driver of autoimmunity. We previously demonstrated an association between MG disease severity and pathogenic Th17 cell frequencies. However, the molecular signatures that differentiate pathogenic and non-pathogenic Th17 cells remain undefined.

Objective To investigate gene signatures and related cytokine profiles associated with pathogenic and non-pathogenic Th17 cells in myasthenia gravis (MG).

Design/Methods Single-cell RNA sequencing (scRNA-seq) analysis was performed on PBMCs from 3 MG patients with high frequencies of pathogenic Th17 cells. PBMCs were differentiated under Th17 polarizing conditions and activated (OX40+CD25+) CD4 T cells were flow-sorted prior to scRNA-seq. To validate the upregulated cytokine transcriptomes from scRNA-seq, supernatants from the enriched CD4 T cell cultures (N=40) were tested with a multiplex cytokine assay.

Results Using scRNA-seq, we identified single CD4 T cells with four cytokine profiles: IFN-γ+IL-17+, IFN-γ+IL-17-, IFN-γ-IL-17+, and IFN-γ-IL-17-. Compared with the other subsets, pathogenic Th17 cells demonstrated significant upregulation of genes associated with cytolysis (GZMK, GZMA), inflammation (IL-9, IL-22), and chemotaxis (CXCL3, CCL4, CCL5). Additionally, pathogenic Th17 cells exhibited higher levels of activation and proliferative capacity. Pseudo-time trajectory analysis demonstrated that pathogenic Th17 cells highly upregulated expression of IL-23R and originated from non-pathogenic Th17 cells. In addition, Granzyme A and IL-17A concentrations in supernatants were significantly higher in severe MG patients than mild patients. IFN-g, IL-9 and MMP-1 concentrations also tended to be higher in severe patients.

Conclusions This investigation shows that Th17 cells in human MG exhibit plasticity and that pathogenic Th17 cells have a unique pro-inflammatory gene signature compared to non-pathogenic Th17 cells. Th17 cells from more severe MG patients also have higher pro-inflammatory cytokines than mild patients. Future investigation is needed to determine whether the unique pathogenic Th17 cell signature could serve as a disease biomarker or target for future therapeutics.

Authors/Disclosures
Yingkai Li, MD, PhD (Duke University) PRESENTER	Dr. Li has nothing to disclose.
	No disclosure on file
Changxin Wan	Mr. Wan has nothing to disclose.
	No disclosure on file
Tabitha Karatz (Duke University)	No disclosure on file
James F. Howard, Jr., MD, FAAN (The University of North Carolina, Dept of Neurology, CB 7025)	Dr. Howard has received personal compensation in the range of $10,000-$49,999 for serving on a Scientific Advisory or Data Safety Monitoring board for arenx . Dr. Howard has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Regeneron Pharmaceuticals. Dr. Howard has received personal compensation in the range of $10,000-$49,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Alexion Pharmaceuticals. Dr. Howard has received personal compensation in the range of $10,000-$49,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Ra Pharma (now UCB Biosciences). Dr. Howard has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Horizon Therapeutics. Dr. Howard has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Sanofi US. Dr. Howard has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Novartis. Dr. Howard has received personal compensation in the range of $10,000-$49,999 for serving on a Speakers Bureau for Academic CME. Dr. Howard has received personal compensation in the range of $10,000-$49,999 for serving on a Speakers Bureau for PeerView CME. Dr. Howard has received personal compensation in the range of $10,000-$49,999 for serving on a Speakers Bureau for Platform Q CME. Dr. Howard has received personal compensation in the range of $5,000-$9,999 for serving on a Speakers Bureau for Zai Laboratories. Dr. Howard has received personal compensation in the range of $10,000-$49,999 for serving on a Speakers Bureau for Medscape CME. Dr. Howard has stock in General Electric dividends. Dr. Howard has stock in Johnson & Johnson dividends. Dr. Howard has stock in Pfizer dividends. An immediate family member of Dr. Howard has stock in GlaxoSmithKline dividends. An immediate family member of Dr. Howard has stock in GE Healthcare dividends. The institution of Dr. Howard has received research support from Alexion Pharmaceuticals. The institution of Dr. Howard has received research support from argenx . The institution of Dr. Howard has received research support from UCB Biosciences. The institution of Dr. Howard has received research support from NIH. The institution of Dr. Howard has received research support from Centers for Disease Control/Research Triangle Institute. The institution of Dr. Howard has received research support from Duke University (DCRI). The institution of Dr. Howard has received research support from Cartestian Therapeutics. Dr. Howard has a non-compensated relationship as a Scientific Advisiory Board member, Committee member with Myasthenia Gravis Foundation of America that is relevant to AAN interests or activities. Dr. Howard has a non-compensated relationship as a Committee member with American Assoc Neuromuscular and Electrodiagnostic Medicine that is relevant to AAN interests or activities.
Janice M. Massey, MD, FAAN (Duke University Medical Center)	Dr. Massey has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Argenix. Dr. Massey has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Revance Therapeutics. Dr. Massey has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Momenta. The institution of Dr. Massey has received research support from Revance .
Vern C. Juel, MD, FAAN (Duke University Medical Center)	Dr. Juel has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Immunovant. Dr. Juel has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Alexion. Dr. Juel has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Accordant Health Services. Dr. Juel has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Janssen. The institution of Dr. Juel has received research support from argenx. The institution of Dr. Juel has received research support from Alexion. The institution of Dr. Juel has received research support from Janssen. The institution of Dr. Juel has received research support from NIH Rare Diseases Network.
Lisa Hobson-Webb, MD, FAAN (Duke University Medical Center)	An immediate family member of Dr. Hobson-Webb has received personal compensation for serving as an employee of G1 Therapeutics. Dr. Hobson-Webb has received personal compensation in the range of $10,000-$49,999 for serving as an Editor, Associate Editor, or Editorial Advisory Board Member for Wiley Publishing. The institution of Dr. Hobson-Webb has received research support from Sanofi Genzyme.
Karissa Gable, MD	Dr. Gable has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Medscape, Web MD. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Pharmacy Times. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Cowen. Dr. Gable has received personal compensation in the range of $10,000-$49,999 for serving as a Consultant for Accordant. Dr. Gable has received personal compensation in the range of $5,000-$9,999 for serving as a Consultant for Immunovant. Dr. Gable has received personal compensation in the range of $10,000-$49,999 for serving as a Consultant for Takeda. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Sanofi. Dr. Gable has received personal compensation in the range of $5,000-$9,999 for serving as a Consultant for Argenx. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Astrazenca. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Guidepoint. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Argenx. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Ultragenx. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Immunovant. Dr. Gable has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Takeda. Dr. Gable has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Sanofi. Dr. Gable has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Argenx. Dr. Gable has received personal compensation in the range of $5,000-$9,999 for serving as a Lecture series with Medscape.
	No disclosure on file
Shruti M. Raja, MD (Duke University)	Dr. Raja has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Signant Health. Dr. Raja has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Regeneron. The institution of Dr. Raja has received research support from American Brain Foundation/Myasthenia Gravis Foundation of America Clinician Scientist Development Award.
Jeff Guptill, MD, FAAN (argenx US)	Dr. Guptill has received personal compensation for serving as an employee of argenx. Dr. Guptill has stock in argenx.