{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T06:04:22Z","timestamp":1789625062139,"version":"build-2803163510"},"posted":{"date-parts":[[2026,9,14]]},"group-title":"Cancer Biology","reference-count":43,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T00:00:00Z","timestamp":1789344000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Cancer Institute","award":["U01 CA272257"],"award-info":[{"award-number":["U01 CA272257"]}]},{"name":"National Cancer Institute","award":["P30 CA77598"],"award-info":[{"award-number":["P30 CA77598"]}]},{"award":["UM1 TR004405"],"award-info":[{"award-number":["UM1 TR004405"]}],"id":[{"id":"https:\/\/ror.org\/04pw6fb54","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Diffuse B-cell lymphomas are the most prevalent canine hematologic malignancies, and their clinical presentation resembles that of human B-cell non-Hodgkin lymphomas (NHLs). However, a lack of a clear subtyping framework perpetuates imprecise treatment approaches that fail to address mechanisms of therapy resistance. Here, we employed deep phenotyping via serial sampling and longitudinal transcriptomics to evaluate intratumoral composition and therapy response in a 6-year-old neutered male goldendoodle with stage 5A marginal zone lymphoma (MZL). Following sequential treatment consisting of a single IV dose of oncolytic vesicular stomatitis virotherapy (VSV) and standard CHOP chemotherapy 30 days later, we performed scRNAseq on seven serial lymph node biopsies and spatial sequencing on a resection taken 10 days post-VSV treatment. B cells (&gt;90%) comprised three transcriptionally distinct and temporally stable subpopulations, with specific copy number profiles and robust spatial organization despite disrupted lymph node architecture. Analysis across subpopulations revealed downregulation of the quiescence program including\n                  <jats:italic>KLF2<\/jats:italic>\n                  , suggesting similarity to\n                  <jats:italic>KLF2<\/jats:italic>\n                  -deficient human MZL. While VSV successfully reached target B cells, downstream transcriptional responses were predominantly localized to the T-cell compartment. T cells (\u22646%) showed increased proliferation and upregulation of cytotoxicity markers post-VSV administration, enriching for leukocyte-mediated cytotoxicity pathways consistent with an anti-viral immune response. Ultimately, these results provide an\n                  <jats:italic>in vivo<\/jats:italic>\n                  proof-of-concept for the treatment paradigm in canine lymphoma, while emphasizing the need for improved subtyping frameworks and multidimensional treatment strategies targeting intratumoral heterogeneity.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance<\/jats:title>\n                  <jats:p>\n                    Using longitudinal single-cell and spatial transcriptomics, we mapped the molecular architecture and therapeutic response of canine marginal zone lymphoma\n                    <jats:italic>in vivo<\/jats:italic>\n                    . We identified conserved\n                    <jats:italic>KLF2<\/jats:italic>\n                    downregulation linking canine and human MZL, alongside stable tumor heterogeneity and a distinct immune response to oncolytic virotherapy. These results demonstrate the utility of deep serial phenotyping to track therapy response, while validating naturally occurring canine cancers as comparative models for human therapeutic development.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.10.750253","type":"posted-content","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T13:50:12Z","timestamp":1789393812000},"source":"Crossref","is-referenced-by-count":0,"title":["Longitudinal single-cell and spatial transcriptomics reveals intratumor heterogeneity, therapeutic response, and comparative value of canine marginal zone lymphoma"],"prefix":"10.64898","author":[{"given":"GE","family":"Walker","sequence":"first","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA"},{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M","family":"Macchietto","sequence":"additional","affiliation":[{"name":"Research Informatics, Minnesota Supercomputing Institute, University of Minnesota, Minneapolis, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K","family":"Reid","sequence":"additional","affiliation":[{"name":"Clinical Investigation Center, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"LE","family":"Burt","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA"},{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Winter","sequence":"additional","affiliation":[{"name":"Clinical Investigation Center, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S","family":"Pracht","sequence":"additional","affiliation":[{"name":"Clinical Investigation Center, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M","family":"Buettner","sequence":"additional","affiliation":[{"name":"Clinical Investigation Center, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V","family":"Penza","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine & Clinical Translation, Mayo Clinic, Rochester, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C","family":"Yung","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R","family":"Dicovitsky","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Kuzmik","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"DA","family":"Vallera","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA"},{"name":"Department of Radiation Oncology, Medical School, University of Minnesota, Minneapolis, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K","family":"Demos-Davies","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"DM","family":"Seelig","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA"},{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Borgatti","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA"},{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M","family":"Henson","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA"},{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C","family":"Feiock","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"},{"name":"Clinical Investigation Center, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"JF","family":"Modiano","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA"},{"name":"Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S","family":"Naik","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine & Clinical 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