Tumor loss-of-function mutations in STK11/LKB1 induce cachexia
Puneeth Iyengar, Aakash Y. Gandhi, Jorge Granados, Tong Guo, Arun Gupta, Jinhai Yu, Ernesto M. Llano, Faya Zhang, Ang Gao, Asha Kandathil, Dorothy Williams, Boning Gao, Luc Girard, Venkat S. Malladi, John M. Shelton, Bret M. Evers, Raquibul Hannan, Chul Ahn, John D. Minna, Rodney E. Infante
Puneeth Iyengar, Aakash Y. Gandhi, Jorge Granados, Tong Guo, Arun Gupta, Jinhai Yu, Ernesto M. Llano, Faya Zhang, Ang Gao, Asha Kandathil, Dorothy Williams, Boning Gao, Luc Girard, Venkat S. Malladi, John M. Shelton, Bret M. Evers, Raquibul Hannan, Chul Ahn, John D. Minna, Rodney E. Infante
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Research Article Metabolism Oncology

Tumor loss-of-function mutations in STK11/LKB1 induce cachexia

Abstract

Cancer cachexia (CC), a wasting syndrome of muscle and adipose tissue resulting in weight loss, is observed in 50% of patients with solid tumors. Management of CC is limited by the absence of biomarkers and knowledge of molecules that drive its phenotype. To identify such molecules, we injected 54 human non–small cell lung cancer (NSCLC) lines into immunodeficient mice, 17 of which produced an unambiguous phenotype of cachexia or non-cachexia. Whole-exome sequencing revealed that 8 of 10 cachexia lines, but none of the non-cachexia lines, possessed mutations in serine/threonine kinase 11 (STK11/LKB1), a regulator of nutrient sensor AMPK. Silencing of STK11/LKB1 in human NSCLC and murine colorectal carcinoma lines conferred a cachexia phenotype after cell transplantation into immunodeficient (human NSCLC) and immunocompetent (murine colorectal carcinoma) models. This host wasting was associated with an alteration in the immune cell repertoire of the tumor microenvironments that led to increases in local mRNA expression and serum levels of CC-associated cytokines. Mutational analysis of circulating tumor DNA from patients with NSCLC identified 89% concordance between STK11/LKB1 mutations and weight loss at cancer diagnosis. The current data provide evidence that tumor STK11/LKB1 loss of function is a driver of CC, simultaneously serving as a genetic biomarker for this wasting syndrome.

Authors

Puneeth Iyengar, Aakash Y. Gandhi, Jorge Granados, Tong Guo, Arun Gupta, Jinhai Yu, Ernesto M. Llano, Faya Zhang, Ang Gao, Asha Kandathil, Dorothy Williams, Boning Gao, Luc Girard, Venkat S. Malladi, John M. Shelton, Bret M. Evers, Raquibul Hannan, Chul Ahn, John D. Minna, Rodney E. Infante

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Figure 5

STK11/LKB1 silencing in a non–cachexia-inducing murine CRC line.

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STK11/LKB1 silencing in a non–cachexia-inducing murine CRC line. (A–M) ...
(A–M) Chow-fed C57BL/6J mice (12-week-old male mice, n = 6–8 per cohort) were injected s.c. with vehicle (black closed circles) or cells from the parental MC38 (black open circles), MC38ΔGFP (blue circles), or 2 different MC38ΔSTK11 (red closed and open circles) lines as described in Methods. Longitudinal measurements of tumor volume (A and F), daily food intake (B), tumor-free body weight at sacrifice (C), tumor-free lean mass at sacrifice (D), and fat mass (E and F) were obtained as described in Methods. Tumor or serum samples at sacrifice were processed for measurement of the levels of tumor mRNA levels relative to MC38 parental cohort of the indicated genes normalized to β-actin (G–K) or serum cytokine levels (L) as described in Methods. Injected cells (A, inset), tumors at sacrifice (A, inset), and epididymal/gonadal white adipose tissue at sacrifice (M) were processed for immunoblot analysis with the indicated antibodies as described in Methods. Data are shown as mean ± SEM of the actual measurement (A and B) or relative to their day 0 values (C–F). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 based on 2-way (A and E) or 1-way (B–D and F–L) ANOVA followed by a Tukey’s (A and E), Dunnett’s (B–D and F–K), or Bonferroni’s (L) multiple-comparison test for significant differences from the MC38ΔSTK11(1) cohort. (N–P) Chow-fed C57BL/6J mice, 12- (n = 8), 14- (n = 8), and 18- (n = 6) week-old males, were injected s.c. with cells from the parental MC38 (black open circles), MC38ΔGFP (blue circles), or MC38ΔSTK11 (red closed and open circles) lines as described in Methods. Longitudinal measurements of tumor volume and fat mass were plotted for each cohort. Linear regression analysis was conducted to determine the association of tumor size and fat mass. Data are shown as dot plots with regression line and 95% confidence band.