Data for: Cytosolic bacterial pathogens activate TLR pathways in tumors that synergistically enhance STING agonist cancer therapies
Data files
May 06, 2025 version files 236.59 KB
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Cell_Press_Data_Record_For_submission.xlsx
232.23 KB
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README.md
4.36 KB
Abstract
Bacterial pathogens that invade the eukaryotic cytosol are distinctive tools for fighting cancer, as they preferentially target tumors and can deliver cancer antigens to MHC-I. Cytosolic bacterial pathogens have undergone extensive preclinical development and human clinical trials, yet the molecular mechanisms by which they activate innate immunity in tumors are unclear. We report that intratumoral delivery of phylogenetically distinct cytosolic pathogens, including Listeria, Rickettsia, and Burkholderia species, elicited anti-tumor responses in established, poorly immunogenic melanoma and lymphoma in mice. We were surprised to observe that although the bacteria required entry to the cytosol, the anti-tumor responses were largely independent of the cytosolic sensors cGAS and STING and instead required TLR signaling. Combining pathogens with STING agonists elicited profound, synergistic anti-tumor effects with complete responses in >80% of mice after a single dose. Small-molecule TLR agonists also synergistically enhanced the anti-tumor activity of STING agonists. The anti-tumor effects required RAG2 but not interferon signaling. Mice cured from the combination therapy developed immunity to cancer rechallenge that required CD8+ T cells. Together, these data provide a framework for enhancing the efficacy of microbial cancer therapies and small-molecule innate immune agonists, via the co-activation of STING and TLRs.
https://doi.org/10.5061/dryad.n2z34tn5z
Description of the data and file structure
Figures in this manuscript fall into 3 categories: 1) tumor volume, 2) mouse survival, and 3) type I interferon (IFN-I) production.
1) Tumor volume: Figures 1A, 1B, 1C, 1D, 1E, 2A, 2B, 3A-3D, 3F-3J, 4A, 4B, 4D, 4E, 5A-F, 6B, 7A
The way to interpret these data is that the Y axis is tumor volume, and the X axis is time. Each column of data is an individual mouse. Mice were injected with the anti-cancer agents on day 0, when tumor volumes measured approximately 6 x 6 x 2.5 mm in each direction. Tumor volumes are reported as ellipsoids using the formula: V = (π/6)ABC. For any days not measured, tumor volumes were averaged between days of measurement. If animals were euthanized, tumor volumes were measured until the last mouse from each group was euthanized. n/a signifies that the data are unavailable because the animal was euthanized.
2) Mouse survival: Figures 1A, 1B, 1C, 2A, 2B, 3E, 4C, 4F, 5G, 5H, 5I, 7B, 8A, 8B
Mice were euthanized when tumor volumes reached 15 mm in any one direction. These data report how long mice survived until their euthanasia. A 1 represents that the animal was euthanized, and a 0 means that the animal survived the entirety of the experiment to the day listed. n/a signifies that the data are unavailable because the animal was euthanized.
3) IFN-I production: Fig. 6A.
IFN-I is a pro-inflammatory cytokine produced downstream of innate immune sensors including STING. IFN-I was measured after infection or treatment of macrophages, then supernatants were collected and used to treat an IFN-I responsive cell line. 4 h later these cells were lysed and relative light units (RLU) were measured after adding a luciferin reagent. Data are expressed as fold change over uninfected.
The titles for each Figure are below:
Figure 1: Intratumoral delivery of cytosolic bacterial pathogens elicits dose-dependent anti-tumor responses in multiple non-immunogenic murine tumor models
Figure 2: Cytosolic access of bacteria promotes the anti-tumor response
Figure 3: The microbe-mediated anti-tumor effects are largely independent of cGAS/STING but require TLR signaling
Figure 4: RAG2 is required for the microbe-mediated anti-tumor effects, while IFNAR or IFNGR are largely dispensable
Figure 5: The anti-tumor effects of bacterial pathogens synergize with the STING agonist S100
Figure 6: IFN-I induction by STING agonists is maintained in infected cells in vitro, and IFNs are not critical for the anti-tumor response in vivo
Figure 7: Small molecule STING+TLR agonist combinations elicit synergistic anti-tumor effects
Figure 8: Bacterial cancer therapies elicit immunity to tumor cell rechallenge
Variable data and abbreviations:
Files and variables
File: Cell_Press_Data_Record_For_submission.xlsx
Description: All tumor volume and survival data, Danielson et al. 2024
Variables
- All reported data of tumor volumes are in cubic millimeters (mm^3).
- Each column is an individual mouse for tumor volume data.
- All survival data is reported as a "1" for the day that an animal was euthanized or a "0" if the animal survived
- Data that are "n/a" means that the mouse was euthanized and therefore no further tumor volumes were collected.
- All drugs/bacteria were administered at day = 0, unless otherwise specified.
- Tumor volume was measured over time and is expressed as an ellipsoid.
- S100, a STING agonist
- The number of bacteria delivered is indicated in log format, for example, "10^7" is 10,000,000.
- Naive means that mice were not immunized.
Abbreviations:
Rp or R. parkeri - Rickettsia parkeri
Lm or L. monocytogenes - Listeria monocytogenes
Bt or B. thailandensis - Burkholderia thailandends
PBS - phosphate-buffered saline
CDN - cyclic dinucleotide (activator of the protein STING)
PAM = PAM3CSK4, a TLR2 agonist
IFNAR = type I interferon receptor
IFNGR = type II / interferon gamma receptor
cGAS = cyclic GMP AMP synthase
Any other abbreviations will be denoted in the manuscript file.
Code/software
NA
Access information
NA
Tumors were injected on the day that they measured approximately 6 x 6 x 2.5 mm in each direction. Tumor volumes are reported as ellipsoids using the formula: V = (π/6)ABC. For any days not measured, tumor volumes were averaged between days of measurement. If animals were euthanized, tumor volumes were measured until the last mouse from each group was euthanized.