Longitudinal single-cell and spatial transcriptomics reveals intratumor heterogeneity, therapeutic response, and comparative value of canine marginal zone lymphoma
- 1University of Minnesota
Abstract
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 (>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 KLF2, suggesting similarity to KLF2-deficient human MZL. While VSV successfully reached target B cells, downstream transcriptional responses were predominantly localized to the T-cell compartment. T cells ([≤]6%) 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 in vivo 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.
Related
- Mutation Order and Selection Shape Intratumor Heterogeneity in Tumor Evolution
- Single-cell transcriptomics-based analysis of cellular heterogeneity in aggrephagy in gastric cancer and development of a prognostic model
- Mapping intestinal microenvironment heterogeneity in Hirschsprung's disease by spatial transcriptomics: Role of THBS1 in macrophage polarization and neural stem cell regulation.
- Spatial Transcriptomics Reveals HER2/AR-Associated Heterogeneity and Alternative Surface Targets in Salivary Duct Carcinoma
- Spatial transcriptomics reveals site-specific cellular and metabolic heterogeneity in bladder carcinoma in situ
BibTeX
@misc{walker2026longitudinal,
title = {{Longitudinal single-cell and spatial transcriptomics reveals intratumor heterogeneity, therapeutic response, and comparative value of canine marginal zone lymphoma}},
author = {G. E. Walker and M. Macchietto and K. Reid and L. E. Burt and A. Winter and S. Pracht and M. Buettner and V. Penza and C. Yung and R. Dicovitsky and A. Kuzmik and D. A. Vallera and K. Demos-Davies and D. M. Seelig and A. Borgatti and M. Henson and C. Feiock and J. F. Modiano and S. Naik and A. L. Sarver and A. E. Treeful},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.10.750253},
url = {https://www.biorxiv.org/content/10.64898/2026.09.10.750253v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Longitudinal single-cell and spatial transcriptomics reveals intratumor heterogeneity, therapeutic response, and comparative value of canine marginal zone lymphoma
AU - G. E. Walker
AU - M. Macchietto
AU - K. Reid
AU - L. E. Burt
AU - A. Winter
AU - S. Pracht
AU - M. Buettner
AU - V. Penza
AU - C. Yung
AU - R. Dicovitsky
AU - A. Kuzmik
AU - D. A. Vallera
AU - K. Demos-Davies
AU - D. M. Seelig
AU - A. Borgatti
AU - M. Henson
AU - C. Feiock
AU - J. F. Modiano
AU - S. Naik
AU - A. L. Sarver
AU - A. E. Treeful
PY - 2026
DA - 2026/09/10
PB - bioRxiv
DO - 10.64898/2026.09.10.750253
UR - https://www.biorxiv.org/content/10.64898/2026.09.10.750253v1
AB - 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 (>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 KLF2, suggesting similarity to KLF2-deficient human MZL. While VSV successfully reached target B cells, downstream transcriptional responses were predominantly localized to the T-cell compartment. T cells ([≤]6%) 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 in vivo 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.
ER -