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| Status |
Public on Jul 05, 2024 |
| Title |
Single Cell RNA sequencing of the non-tumor components of the tumor mass after either empty vector control or Zbtb46 overexpressing lentiviral intratumoral treatments |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Tumor-angiogenesis and -immunity play critical roles in cancer progression and outcome. An inverse correlation of these two1 hints at common regulatory mechanism(s). Here, we report that Zbtb46, a repressive transcription factor and a widely accepted marker for classical dendritic cells (DCs)2,3, constitutes one such regulatory mechanism. Zbtb46 was downregulated in both DCs and endothelial cells (ECs) by tumor-derived factors to facilitate robust tumor growth. Zbtb46 downregulation led to a hallmark pro-tumor microenvironment (TME), including dysfunctional vasculature and immunosuppressive cell accumulation. Analysis of cancer patient data revealed a similar association of low ZBTB46 expression with an immunosuppressive TME and a worse prognosis. In contrast, enforced Zbtb46 expression brought dynamic changes in the TME landscape to mitigate the pro-tumor features and to enhance anti-tumor immune components, restricting tumor growth. Mechanistically, Zbtb46-deficient ECs were highly angiogenic, and Zbtb46-deficient bone-marrow progenitors upregulated Cebpb and diverted the DC program to myeloid lineage output, potentially explaining the myeloid lineage skewing phenomenon in cancer4. Conversely, enforced Zbtb46 expression normalized tumor vessels and, by suppressing Cebpb, skewed bone-marrow precursors towards more DC generation over macrophages, leading to an immune-hot TME. Remarkably, Zbtb46 mRNA treatment synergized with anti-PD1 immunotherapy to improve tumor management in pre-clinical models. These findings identify Zbtb46 as a common regulatory mechanism for angiogenesis and for myeloid lineage skewing in cancer and suggest that maintaining its expression could have therapeutic benefits.
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| Overall design |
scRNA sequencing of the non-tumor components of the PyMT-BO1 bearing mice on day 16 after either empty vector control or Zbtb46 overexpressing lentiviral intratumoral treatments on Days 9, 12, and 15 post-tumor-transplantation were performed. Briefly, on day 16, tumor masses from a total of 5 mice were pulled in each group. Following enzymatic digestion, the cells were FACS sorted as live non-tumor (GFP-) population. For multiplexing, cells were split into separate tubes as either mCherry+ or mCherry- cells (mCherry is a fluorescence reporter included in both the empty vector and Zbtb46 overexpressing vector), marked with distinctive hashtags (Hashtag-B0301 for mCherry+ and Hashtag-B0302 for mCherry- cells), and pulled back to make two samples: control and treatment. The cells were then processed for single-cell library generation, using the 10X Genomics platform, and sequenced with Illumina NovaSeq 6000.
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| Contributor(s) |
Kabir AU, Zeng C, Artyomov M, Choi K |
| Citation missing |
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| Submission date |
Apr 16, 2024 |
| Last update date |
Jul 06, 2024 |
| Contact name |
Ashraf Ul Kabir |
| E-mail(s) |
a.kabir@wustl.edu
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| Organization name |
Washington University in St. Louis
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| Department |
Pathology and Immunology
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| Street address |
660 S Euclid Ave, BJCIH 8th floor, room 8302
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| City |
St. Louis |
| State/province |
Missouri |
| ZIP/Postal code |
63110-1093 |
| Country |
USA |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA1101021 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE264124_RAW.tar |
908.5 Mb |
(http)(custom) |
TAR (of MTX, TSV) |
SRA Run Selector |
| Raw data are available in SRA |
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