NPRL2 gene therapy induces effective antitumor immunity in KRAS/STK11 mutant anti-PD1 resistant metastatic non-small cell lung cancer (NSCLC) in a humanized mouse model
- Department of Thoracic and Cardiovascular Surgery, The University of Texas MD Anderson Cancer Center, Houston, United States
- Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, United States
- Department of Stem Cell Transplantation, The University of Texas MD Anderson Cancer Center, Houston, United States
Figures
Figure 1 with 1 supplement
Reduced expression of NPRL2 in non-small cell lung cancer (NSCLC) and its restoration caused inhibition of colony formation and apoptosis induction.
(A) Basal level of NPRL2 and its correspondent death protein ligand 1 (PD-L1) expression were evaluated on anti-PD1 resistant KRAS/LKB1 mutant A549 and sensitive H1299 cells. (B) Inhibition of …
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Figure 1—source data 1
Reduced expression of NPRL2 in non-small cell lung cancer (NSCLC) and its restoration caused inhibition of colony formation and apoptosis induction.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig1-data1-v1.xlsx
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Figure 1—source data 2
PDF containing original western blots for Figure 1A, indicating the relevant bands and samples.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig1-data2-v1.pdf
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Figure 1—source data 3
Original files for western blot images displayed in Figure 1A.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig1-data3-v1.zip
Figure 1—figure supplement 1
Basal level of NPRL2 expression and its transfection on various NSCLC cell lines.
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Figure 1—figure supplement 1—source data 1
PDF file containing original western blots for Figure 1—figure supplement 1, indicating the NPRL2 expressions in different NSCLC cell lines.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig1-figsupp1-data1-v1.pdf
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Figure 1—figure supplement 1—source data 2
Original files for western blot images displayed in Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig1-figsupp1-data2-v1.zip
Figure 2
Antitumor effect of NPRL2 gene therapy on anti-PD1 resistant KRAS/LKB1 mutant tumors in humanized and non-humanized mice.
(A) Experimental strategy for non-humanized NSG mice shows that tumors were developed through intravenous injection of anti-PD1 resistant A549-luc cells and lung metastasis were treated with NPRL2 …
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Figure 2—source data 1
Antitumor effect of NPRL2 gene therapy on anti-PD1 resistant KRAS/LKB1 mutant tumors in humanized and non-humanized mice.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig2-data1-v1.xlsx
Figure 3
NPRL2 induces antitumor immune responses in anti-PD1 resistant KRAS/LKB1 mutant A549-Lung Met in humanized mice.
A549 lung metastases was developed in 6–8 wk post-humanized mice and lung met-bearing humanized mice were then treated with NPRL2, pembrolizumab, and its combination. 3-to 5 d after the treatment, …
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Figure 3—source data 1
NPRL2 induces antitumor immune responses in anti-PD1 resistant KRAS/LKB1 mutant A549-Lung Met in humanized mice: Tumor microenvironment analysis in humanized mice.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig3-data1-v1.xlsx
Figure 4
NPRL2 induced synergistic antitumor immune response with pembrolizumab on anti-PD1 responsive H1299 tumors in humanized mice.
(A) Experimental strategy of H1299 subcutaneous tumor development in humanized mice. NSG mice were humanized for 7–8 wk and humanization was verified by blood screening followed by tumor cell …
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Figure 4—source data 1
NPRL2 induced synergistic antitumor immune response with pembrolizumab on anti-PD1 responsive H1299 tumors in humanized mice: NPRL2 antitumor effect on H1299 tumors and tumor microenvironment analysis.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig4-data1-v1.xlsx
Figure 5
Antitumor effect of NPRL2 on anti-PD1 resistant LLC2 tumors in a syngeneic mouse model.
(A) Treatment strategy of anti-PD1 resistant LLC tumors. Subcutaneous tumors were developed in C57BL/6 mice followed by NPRL2 (i.v.) 25 μg/mouse and anti-PD1 treatments for 3 wk. Tumor volume was …
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Figure 5—source data 1
Antitumor effect of NPRL2 on anti-PD1 resistant LLC2 tumors in a syngeneic mouse model and their tumor microenvironment analysis.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig5-data1-v1.xlsx
Figure 6
Antitumor effect of NPRL2 on anti-PD1 resistant LLC2 tumors after depletion of immune cells in a syngeneic mouse model.
(A) Treatment strategy for immune cell depletion experiment. Antibody-mediated CD4 + T, CD8 + T, natural killer (NK) cell depletion, and clodronate-mediated macrophage and dendritic cell depletion …
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Figure 6—source data 1
Antitumor effect of NPRL2 on anti-PD1 resistant LLC2 tumors in immune cells depleted syngeneic mice.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig6-data1-v1.xlsx
Figure 7
Alteration of gene expression and immune signaling associated with T cell activation by NPRL2 in humanized mice.
Expression of 770 human immune-related genes was determined in anti-PD1 resistant lung metastasis in humanized mice by nano string and the effect of NPRL2 on gene expression was analyzed. (A) A …
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Figure 7—source data 1
Alteration of gene expression and immune signaling associated with T cell activation by NPRL2 in humanized mice: Nano string analysis in tumors developed in humanized mice.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig7-data1-v1.csv
Figure 8 with 1 supplement
Restoration of NPRL2 expression altered tumor microenvironment (TME), induced apoptosis, inhibited cell growth, and signaling.
NPRL2 stable clones in A549 and H1299 NSCLC cells were generated and developed tumors in humanized mice for tumor microenvironment analysis. The in-vitro assays were performed using these stable …
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Figure 8—source data 1
Restoration of NPRL2 expression altered tumor microenvironment (TME), induced apoptosis, inhibited cell growth and signaling: Tumor microenvironment analysis in NPRL2 stable expressing tumors.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig8-data1-v1.xlsx
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Figure 8—source data 2
PDF file containing original western blots for Figure 8R and S, indicating relevant bands, treatments and samples.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig8-data2-v1.pdf
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Figure 8—source data 3
Original files for western blot images displayed in Figure 8R and S.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig8-data3-v1.zip
Figure 8—figure supplement 1
Stable expression NPRL2 affected the cell cycle.
H1299 parental and NPRL2 stably expressing cells were subjected to cell cycle assay. Upper panel showed the different phases such as G1, S, and G2/M in both H1299 and H1299-NPRL2++/++ cells. Bottom …
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Figure 8—figure supplement 1—source data 1
Cell cycle analysis in NPRL2 stably expressing cells.
- https://cdn.elifesciences.org/articles/98258/elife-98258-fig8-figsupp1-data1-v1.xlsx
