American Association for Cancer Research
00085472can181207-sup-200471_2_supp_5130234_ph6rqw.doc (1.8 MB)

Supplementary Methods, Figure S1, Figure S2, Figure S3, Figure S4, Figure S5, Figure S6, Figure S7, Figure S8, Table S1, Table S2, Table S3, Table S4 from Mas Receptor Activation Slows Tumor Growth and Attenuates Muscle Wasting in Cancer

Download (1.8 MB)
journal contribution
posted on 2023-03-31, 02:45 authored by Kate T. Murphy, Mohammed I. Hossain, Kristy Swiderski, Annabel Chee, Timur Naim, Jennifer Trieu, Vanessa Haynes, Suzannah J. Read, David I. Stapleton, Sarah M. Judge, Jose G. Trevino, Andrew R. Judge, Gordon S. Lynch

This file contains: • Supplemental Methods • Supplemental Figure S1. Muscle-specific overexpression of the AT1 but not the MasR regulates skeletal muscle fiber size in healthy mice. • Supplemental Figure S2. Representative images of differentiated C2C12 myotubes incubated for 48 h in normal horse serum and treated with vehicle control or the ACE/Ang II/AT1 axis activators Ang II or A779 • Supplemental Figure S3. Genetic MasR overexpression was induced by transducing C2C12 myoblasts with a Tet-regulated MasR lentivirus or empty lentivirus and treating differentiated myotubes with doxycylin • Supplemental Figure S4. Low dose AVE 0991 induces small but significant improvements in food and water intake and weight loss in severely cachectic C-26 tumor-bearing mice • Supplemental Figure S5. Low dose AVE 0991 (1 mg/kg) causes an oxidative-to-glycolytic shift in fiber types in TA muscles from severely cachectic C-26 tumor-bearing mice • Supplemental Figure S6. High dose AVE 0991 treatment enhanced cumulative water intake and movement as assessed by the number of beam breaks in severely cachectic C-26 tumor-bearing mice fed ad libitum but not in those pair-fed to vehicle treated controls • Supplemental Figure S7. Pharmacological Ang-(1-7)/MasR activation protects against Lewis Lung Carcinoma (LLC) cancer cell induced muscle fiber atrophy in vitro • Supplemental Figure S8. Specificity of our custom anti-AT1 peptide compared to a commercially available anti-AT1 antibody • Supplemental Table S1. Primers used to amplify AT1 and MasR genes • Supplemental Table S2. Human cancer cachexia study group characteristics • Supplemental Table S3. List of upregulated proteins in AVE 0991 treated serum starved myotubes • Supplemental Table S4. List of down regulated proteins in AVE 0991 treated serum starved myotubes


Department of Neurology, The University of Washington


National Institute of Arthritis, Musculoskeletal and Skin Diseases



Cancer cachexia is a multifactorial syndrome characterized by a progressive loss of skeletal muscle mass associated with significant functional impairment. Cachexia robs patients of their strength and capacity to perform daily tasks and live independently. Effective treatments are needed urgently. Here, we investigated the therapeutic potential of activating the “alternative” axis of the renin-angiotensin system, involving ACE2, angiotensin-(1-7), and the mitochondrial assembly receptor (MasR), for treating cancer cachexia. Plasmid overexpression of the MasR or pharmacologic angiotensin-(1-7)/MasR activation did not affect healthy muscle fiber size in vitro or in vivo but attenuated atrophy induced by coculture with cancer cells in vitro. In mice with cancer cachexia, the MasR agonist AVE 0991 slowed tumor development, reduced weight loss, improved locomotor activity, and attenuated muscle wasting, with the majority of these effects dependent on the orexigenic and not antitumor properties of AVE 0991. Proteomic profiling and IHC revealed that mechanisms underlying AVE 0991 effects on skeletal muscle involved miR-23a–regulated preservation of the fast, glycolytic fibers. MasR activation is a novel regulator of muscle phenotype, and AVE 0991 has orexigenic, anticachectic, and antitumorigenic effects, identifying it as a promising adjunct therapy for cancer and other serious muscle wasting conditions. These findings demonstrate that MasR activation has multiple benefits of being orexigenic, anticachectic, and antitumorigenic, revealing it as a potential adjunct therapy for cancer.See related commentary by Rupert et al., p. 699

Usage metrics

    Cancer Research