{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T04:39:38Z","timestamp":1793939978055,"version":"4.8.0"},"posted":{"date-parts":[[2025,10,13]]},"group-title":"Genomics","reference-count":135,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T00:00:00Z","timestamp":1760313600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["UG3NS132139"],"award-info":[{"award-number":["UG3NS132139"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R33 CA267219"],"award-info":[{"award-number":["R33 CA267219"]}],"id":[{"id":"https:\/\/ror.org\/040gcmg81","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"NIDDK","doi-asserted-by":"crossref","award":["P30 DK132710"],"award-info":[{"award-number":["P30 DK132710"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"crossref"}]},{"award":["RM1HG011014"],"award-info":[{"award-number":["RM1HG011014"]}],"id":[{"id":"https:\/\/ror.org\/00baak391","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Career Award for Medical Scientists"],"award-info":[{"award-number":["Career Award for Medical Scientists"]}],"id":[{"id":"https:\/\/ror.org\/01d35cw23","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Scholar Award"],"award-info":[{"award-number":["Scholar Award"]}],"id":[{"id":"https:\/\/ror.org\/05nmp3276","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Blood Cancer United"},{"id":[{"id":"https:\/\/ror.org\/00v7th354","id-type":"ROR","asserted-by":"publisher"}]},{"name":"MacMillan Family Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,10,13]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The human somatic genome evolves throughout our lifespan, producing mosaic individuals comprising clones harboring different mutations across tissues. While clonal expansions in the hematopoietic system have been extensively characterized and reported to be nearly ubiquitous, clonal mosaicism (CM) has more recently also been described across multiple solid tissues. However, outstanding questions remain about the parameters and processes of human somatic evolution in non-cancerous solid human tissues, including when clones arise, how they evolve over time, and what mechanisms lead to their expansion. Questions of timing and clonal dynamics can be addressed through phylogenetic reconstruction, which serves as a \u2018temporal microscope\u2019, while uncovering the mechanisms of expansion necessitates simultaneous phenotypic profiling. To address this gap, here we develop Single-cell Miniaturized Automated Reverse Transcription and Primary Template-directed Amplification (SMART-PTA) for joint single-cell whole-genome and whole-transcriptome sequencing for large scale and cost efficient interrogation of solid tissue CM. We established a workflow that generates hundreds of matched single-cell whole genome and transcriptome libraries within a week. We profiled phenotypically normal esophagus tissue from four aged donors and used somatic variants to build high-resolution single-cell lineages from &gt;2,700 cells with accompanying transcriptomic information, reconstructing &gt;70 years of somatic evolution. T cell expansions identified from T cell receptor (TCR) sequences validated the clonal structure of the single-nucleotide variant (SNV)-based phylogenies and phylogenetic cross-correlation analysis showed that epithelial cells had higher degrees of shared ancestry by spatial location compared to immune cells. Mapping mutation signatures to the phylogenetic tree revealed the emergence of tobacco\/alcohol exposure-related signatures later in life, consistent with the donors\u2019 exposure histories. We identified variants in driver genes that were previously reported in the phenotypically normal esophagus, detecting clonal expansions harboring mutations in genes including\n                  <jats:italic>TP53<\/jats:italic>\n                  and\n                  <jats:italic>FAT1<\/jats:italic>\n                  . We mapped the evolution of clones with both monoallelic as well as biallelic\n                  <jats:italic>TP53<\/jats:italic>\n                  loss, including a clone associated with high expression of cell cycling genes and higher chromosome instability. Leveraging the matched transcriptome data, we uncovered cell type biases in mutant clones, with a higher proportion of\n                  <jats:italic>TP53<\/jats:italic>\n                  or\n                  <jats:italic>FAT1<\/jats:italic>\n                  -mutant cells in an earlier basal epithelial cell state compared to wild-type cells. We further observed copy-neutral loss of heterozygosity (CNLOH) events on chromosome 9q that spanned the\n                  <jats:italic>NOTCH1<\/jats:italic>\n                  locus in up to \u223c35% of epithelial cells. Mapping CNLOH events to the phylogenetic tree revealed a striking pattern in which CNLOH was separately acquired many times, reflecting convergent evolution. Cells with CNLOH events were biased towards the earlier basal epithelial state, suggestive of a selective advantage that leads to prevalent recurrence of chr9q CNLOH. Together, we demonstrate that SMART-PTA is an efficient, scalable approach for single-cell whole-genome and whole-transcriptome profiling to build phenotypically annotated single-cell phylogenies with enough throughput and power for application to normal tissue somatic evolution. Moreover, we reconstruct the evolutionary history of the esophageal epithelium at high scale and resolution, providing a window into the dynamics and processes that shape clonal expansions in phenotypically normal tissues throughout a lifespan.\n                <\/jats:p>","DOI":"10.1101\/2025.10.11.681805","type":"posted-content","created":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T04:05:13Z","timestamp":1760414713000},"source":"Crossref","is-referenced-by-count":6,"title":["Large-scale single-cell phylogenetic mapping of clonal evolution in the human aging esophagus"],"prefix":"10.64898","author":[{"given":"Tamara","family":"Prieto","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dennis 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