Abstract 1230: The pre-metastatic niche establishment through the induction of the epithelial-to-mesenchymal transition by circulating exosomes in lymphangioleiomyomatosis (LAM): implication of the complement cascade
Authors: Jouida Amina , McFadden Patrick , Buckley Grace , Lynn Evelyn , Marissa O'Callaghan , Sekimoto Yasuhito , Fabre Aurelie , Keane Michael P. , McCarthy Cormac 
Published: April 2023

Exosomes are a class of extra cellular vesicles (EVs), with a multi vesicular endosomal origin, that are released by all cell types, with sizes ranging from 30-150nm and a lipid bilayer membrane. They have been shown to play key roles in disease progression and have diagnostic and disease monitoring potential. This study aims to characterize exosomes from the serum of patients with Lymphangioleiomyomatosis (LAM), a rare, low-grade, metastasizing neoplasm that occurs predominantly in females. Using Nanoparticle Tracking Analysis (NTA), serum from LAM patients (n=19) have significantly increased numbers of exosomes compared to serum samples from healthy controls (n=20) (8.9x109 vs 13.8x109 particles/ml; p=0.024). Furthermore, particles counts are significantly negatively correlated with the forced expiratory volume (FEV1) (r=-0.4667; p-value=0.0295) in LAM patients. Metalloproteinase activity in exosomes was measured by zymography. Interestingly, these LAM-derived exosomes had significantly higher activity of Pro-MMP-9 (3-fold increase; p-value=0.0278). We analyzed the effects of serum-derived exosomes on cells from an A549 cell line. Increased expression of vimentin, a major marker of EMT, was observed in cells treated with LAM-derived exosomes compared to healthy controls (1.4-fold increase; p=0.0217). Moreover, we showed by a modified Boyden chamber assay that exosomes derived from serum treatment increases the invasive capacity of A549 cells (fold change of 2.2; p-value=0.0004). We interrogated the LAM Single Cell Atlas and identified increased expression of TWIST1, a key transcription factor for EMT. TWIST1 was increased 8-fold in the lung and was uniquely expressed in 'LAM cells', which had metastasized. Furthermore, proteomic analysis revealed high expression of C3 and C5 in LAM-derived exosomes (respectively 1.4- and 2.1-fold change; p=0.0276 and p=0.0034) suggesting the potential regulatory role of the compliment system in the EMT mediated by exosomes These preliminary results highlight the urge to study the specific mechanistic interplay between MMPs and EMT, mediated by exosomes, and whether exosomes are involved in the metastatic spread of LAM cells to the lung and their role in influencing innate and adaptive immune cell response to this invasion.

Citation Format: Amina Jouida, Patrick McFadden, Grace Buckley, Evelyn Lynn, O'Callaghan Marissa, Yasuhito Sekimoto, Aurelie Fabre, Michael P. Keane, Cormac McCarthy. The pre-metastatic niche establishment through the induction of the epithelial-to-mesenchymal transition by circulating exosomes in lymphangioleiomyomatosis (LAM): implication of the complement cascade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1230.