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[Preprint]. 2025 Sep 17:2025.09.14.676175. [Version 1] doi: 10.1101/2025.09.14.676175

Single-cell Genomic Copy Number Evolution Reveals Frequent Loss of the Y chromosome in Esophageal Adenocarcinoma

Shiwei Yin, Na Zhong, Swati Agrawal, Bingru Feng, Sneha Pradhan, Nathan K Jobalia, Maryam Vaziripour, Sampath K Poreddy, Laura A Kresty, Erin A Gillaspie, Nicholas E Dietz, Waddah B Al-Refaie, Kalyana C Nandipati, Yusi Fu, Jun Xia
PMCID: PMC12458328  PMID: 41000889

Abstract

The incidence of esophageal adenocarcinoma (EAC), an aggressive cancer, is rising in Western countries, and incidence is higher in men. To better understand the roles of DNA copy number evolution, tumor heterogeneity, and tumor microenvironment in EAC and its origins, we evaluated EAC, Barrett’s Esophagus (BE), and normal adjacent tissue using our previously developed ultra-sensitive, single-nucleus DNA copy number analysis (12,186 nuclei, 20 specimens) and single-nucleus RNA-sequencing (snRNA-seq; 87,027 nuclei, 32 specimens). Loss of Y (LOY) chromosome was rare in BE but markedly enriched in EAC. In genome unstable cells, complete LOY chromosome often coincided with X-chromosome duplication. We did not detect X-chromosome inactivation via XIST expression in cells with LOY, suggesting increased expression of X-chromosome genes and potential effects on tumor immune evasion. When considering patients’ clinical features, we identified distinct differences in tumor microenvironment by sex, age, and obesity status, particularly massively reduced immune cell composition in obese patients. Finally, our cell line models recapitulate LOY, and we identify the first candidate anti-LOY agent. Our findings highlight how LOY affects tumor subpopulations and shapes the tumor microenvironment during EAC progression.

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