Table 2.

TP53-inactivation induced senescence differentially influences the tumor microenvironment in M0 and M1 tumors. We developed age and sex adjusted regression models using single-sample GSEA (ssGSEA) enrichment scores from TCGA RCC tumors, M0 (n = 297) and M1 (n = 79), to determine whether the TP53-inactivation induced senescence pathway, SENESCENCE_TP53_TARGETS_UP, associates with VHL-induced hypoxia and expression of key immune markers linking cellular senescence and immune infiltration. In models 1–4, we show the association between senescence enrichment and cGAS ($M1$), STING ($M2$), PRF1 $\left(M3\right)$, and GZMA $\left(M4\right)$ expression by regressing SENESCENCE_TP53_TARGETS_UP enrichment scores on the log10 normalized gene expression of these markers. In model 5 $\left(M5\right)$ we show the association between senescence enrichment and hypoxia (MANIA_HYPOXIA_VHL_UP) enrichment in M0 and M1 RCC tumors. Our regression models adjust for age and sex and specifically examine if there is an interaction between SENESCENCE_TP53_TARGETS_UP enrichment and metastasis status in order to determine if tumor senescence activity associates with markers of inflammation differently based on M0 or M1 status.

Model	Outcome	Variable	$\hat{\beta }$	SE	p-value
$M1$	$E\left[g\right]\sim$ cGAS	$E{S}_{senescence\mathrm{\_}UP}$	1.365	0.3475	0.000102
		$E{S}_{senescence\mathrm{\_}UP}\ast M$	−2.714	0.7275	0.000221
$M2$	$E\left[g\right]\sim$STING	$E{S}_{senescence\mathrm{\_}UP}$	1.660	0.2838	1.08*10−08
		$E{S}_{senescence\mathrm{\_}UP}\ast M$	−1.551	0.5942	0.00941
$M3$	$E\left[g\right]\sim$PRF1	$E{S}_{senescence\mathrm{\_}UP}$	2.397	0.5247	6.66*10−06
		$E{S}_{senescence\mathrm{\_}UP}\ast M$	−3.236	1.098	0.00342
$M4$	$E\left[g\right]\sim$GZMA	$E{S}_{senescence\mathrm{\_}UP}$	2.054	0.6228	0.00106
		$E{S}_{senescence\mathrm{\_}UP}\ast M$	−2.930	1.303	0.0252
$M5$	$E\left[p\right]\sim P_{Hypoxia}$	$E{S}_{senescence\mathrm{\_}UP}$	0.7909	0.09713	5.97*10−15
		$E{S}_{senescence\mathrm{\_}UP}\ast M$	−0.5865	0.2033	0.00415

$M1-M4:E\left[g\right]={\hat{\beta }}_{1}E{S}_{senescence\mathrm{\_}UP}+{\hat{\beta }}_{2}M+{\hat{\beta }}_{3}E{S}_{senescence\mathrm{\_}UP}\ast M+{\hat{\beta }}_{4}a+{\hat{\beta }}_{5}s$

$M5:E\left[p\right]={\hat{\beta }}_{1}E{S}_{senescence\mathrm{\_}UP}+{\hat{\beta }}_{2}M+{\hat{\beta }}_{3}E{S}_{senescence\mathrm{\_}UP}\ast M+{\hat{\beta }}_{4}a+{\hat{\beta }}_{5}s$ where$E\left[g\right]$is the expected $lo{g}_{10}$ normalized gene expression of cGAS (MB21D1), STING, PRF1, or GZMA, $E{S}_{senescence\mathrm{\_}UP}$ is SENESCENCE_TP53_TARGETS_UP pathway enrichment scores, $E\left[p\right]$is the expected pathway enrichment score of $P_{Hypoxia}$, the MANIA_HYPOXIA_VHL_TARGETS_UP pathway, $M$ is metastasis status, $a$ is age, and $s$ is sex.

Note: SE is standard error