Host autophagy mediates organ wasting and nutrient mobilization for tumor growth

• DOI: 10.15252/embj.2020107336
• Publication Date: 2021-07-26T10:16:01Z
• Authors: Rojyar Khezri; Petter Holland; Todd Andrew Schoborg; Ifat Abramovich; Szabolcs Takáts; Caroline Dillard; Ashish Jain; Fergal O'Farrell; Sebastian Wolfgan...; William M Hagopian; Eduardo Martin Qu...; Rachel Ng; Nadja Sandra Kath...; Mohammed Mahidur ...; José Gerardo Tele...; Andreas Brech; Heinrich Jasper; Nasser M Rusan; Anne Hope Jahren; Eyal Gottlieb; Tor Erik Rusten
• Keywords: 0301 basic medicine; 570; Cachexia; 610; Articles; Nutrients; 12. Responsible consumption; Disease Models, Animal; 03 medical and health sciences; Drosophila melanogaster; Neoplasms; Autophagy; Disease Progression; Animals; Humans; Energy Metabolism; Muscle, Skeletal

Article26 July 2021Open Access Source DataTransparent process Host autophagy mediates organ wasting and nutrient mobilization for tumor growth Rojyar Khezri Centre Cancer Cell Reprogramming, Institute of Clinical Medicine, Faculty University Oslo, Norway Department Molecular Biology, Research, Oslo Hospital, Search more papers by this author Petter Holland Todd Andrew Schoborg Biology Physiology Center, National Heart, Lung Blood Institute, Institutes Health, Bethesda, MD, USA Ifat Abramovich The Ruth Bruce Rappaport Technion - Israel Technology, Bat Galim, Haifa, Szabolcs Takáts Caroline Dillard Ashish Jain orcid.org/0000-0001-6549-2788 Fergal O'Farrell Sebastian Wolfgang Schultz orcid.org/0000-0002-3661-2178 William M Hagopian Earth Evolution Dynamics, Eduardo Martin Quintana Rachel Ng orcid.org/0000-0001-8969-9628 Nadja Sandra Katheder Immunology Discovery, Genentech, Inc., South San Francisco, CA, Mohammed Mahidur Rahman orcid.org/0000-0001-5327-4193 José Gerardo Teles Reis Andreas Brech Heinrich Jasper Buck Research on Aging, Novato, Nasser Rusan Anne Hope Jahren Eyal Gottlieb orcid.org/0000-0002-9770-0956 Tor Erik Rusten Corresponding Author [email protected] orcid.org/0000-0002-9150-2676 Information Khezri1,2, Holland1,2,†, Schoborg3,†, Abramovich4, Takáts1,2, Dillard1,2, Jain1,2, O'Farrell1,2, Schultz1,2, Hagopian5, Quintana1,2, Ng3, Katheder2,6, Rahman1,2, Reis1,2, Brech1,2, Jasper6,7, Rusan3, Jahren5, Gottlieb4 *,1,2 1Centre 2Department 3Cell 4The 5Centre 6Immunology 7Buck † These authors contributed equally to work *Corresponding author. E-mail: EMBO Journal (2021)40:e107336https://doi.org/10.15252/embj.2020107336 PDFDownload PDF article text main figures. Peer ReviewDownload a summary the editorial decision including letters, reviewer comments responses feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract During growth—when anabolic demands are high—autophagy supports metabolism through lysosomal organelle turnover recycling. Ras-driven tumors additionally invoke non-autonomous in microenvironment support growth, part transfer amino acids. Here we uncover third critical role mediating systemic using well-characterized malignant model Drosophila melanogaster. Micro-computed X-ray tomography metabolic profiling reveal that RasV12; scrib−/− grow 10-fold volume, while unfolds with progressive muscle atrophy, loss body mass, -motility, -feeding, eventually death. Tissue is found be mediated results host acids sugars into circulation. Natural abundance Carbon 13 tracing demonstrates biomass increasingly derived from tissues as source progresses. We conclude utilized growth. Synopsis Autophagy maintains mitochondrial health recycling cells, promotes microenvironmental thereby sustaining In study, tomography, metabolomics carbon autophagy-mediated distal provides increase eye RasV12, induce cause release sugar Systemic release. Tumor derive ensues. Introduction Macroautophagy (referred herein) encapsulates cytoplasm double membrane vesicle subsequently degraded upon fusion lysosome. Through autophagy, cytoplasmic cargo, glycogen, protein aggregates, organelles such mitochondria, broken down reused energy production or macromolecular building blocks (Galluzzi et al, 2015). Physiologically, necessary survive starvation both unicellular organisms animal models. Genetic studies have revealed required sustain circulating acid levels autophagy-deficient newborn mice (Kuma 2004) glucose during fasting adult (Karsli-Uzunbas 2014). As essential cellular control levels, focused effort has been understanding relevance carcinogenesis (Kimmelman White, 2017; Poillet-Perez 2019). Indeed, when high, findings several genetically engineered mouse models uncovered tumor-supportive (Poillet-Perez deficiency KrasG12D BrafV600E lung cancer, BrafV600E;PTEN−/− KrasG12D, pancreatic cancer (PDAC) decreased progression (Guo 2013; Rosenfeldt Strohecker Karsli-Uzunbas 2014; Yang Xie Metabolic analyses RAS-driven cells KrasV12D; LKB1−/− showed cell-autonomously provide substrates TCA cycle maintaining nucleotide pools prevent crisis (Bhatt Thus, functions order promote progression. not however only within cell itself. flies mice, neighboring (Sousa 2016; 2018). Drosophila, also induced stress response adipose tissue, an observation akin effects late-stage patients suffering cachexia syndrome (Katheder 2017). tumor-induced cascade events leads inflammation, reprogramming, degeneration, particularly fat skeletal heart whereas liver increases size (Baracos Muscle samples (Tardif Aversa 2016) Xenograft (Penna 2013) show increased molecular markers autophagy. This led idea may executed intracellular material simultaneous elevated proteasomal autophagic activity experiments whether rate-limiting how affect vivo remain limited. recent knockdown endocytosis regulator BECN1 moderately reduced but morphological changes Results Gradual atrophy weight ensues To assess dynamics at whole-animal level, adopted computed (CT), gold standard evaluating patients. Genetically GFP-labeled invade central nervous system (CNS), extend larval stage kill day 10–12 (Brumby Richardson, 2003; Pagliarini Xu, 2003). optimized fixation staining protocol high-resolution micro-CT (μ-CT) imaging developmentally staged larvae (Schoborg enabled ready identification, segmentation, calculation volumes (Fig 1A–C; Movie EV1–7) 2019) volume invading enveloping brain 6 10 1C, EV7, quantified 2A). Conversely, total initially similar animals carrying benign RasV12 tumors, progressively shrink approximately 50% 1B, 2B). body, which perform functions, displayed striking translucency lipid droplet (Figs 2E 3C) Ras; tumor-carrying 8 (Figueroa-Clarevega Bilder, 2015), although remained unaltered 2D). Organ were accompanied 35% dry motility feeding 2F–H). established can imaged intact heat-killed whole myosin heavy chain-GFP reporter backlight microscopy 3A–C EV1A–C). inhibit ecdysone synthesis offset normal pupation due dilp8 secretion tissue (Colombani 2012; Garelli 2012). No was observed where molting hormone specifically obliterated ecdysoneless (ecd1−/−) mutants, genetic elimination ecdysone-producing prothoracic gland EV1-EV4). onset precede reduction motility, arguing together simple function food intake extended stage. Figure 1. Tumor-induced A. Representative 2D μ-CT scans ctrl 6, 10. (green), (blue) eye-antennal discs/tumor (red) outlined. Scale bar: 1 mm. Anterior (A), Right (R). B. 3D rendering genotypes indicated (A). Dorsal (D), Left (L). C. (yellow), over time. Download figure PowerPoint 2. Quantification (n = 15) 15), 7 5), 4), 9 5). Quantifications 6(n dat width Oblique 3 (DO3) 29) scrib−/−at days 31). D. E. confocal images bearing ages. Lipid droplets highlighted Tox staining. 50 μm. F. 9), 18) tumor-bearing excluding weight. G. measure crawling distance pattern 14), 16), each colored line represents single larva. H. Coomassie assay asses time, three repeated measurements average 20 larvae. Data information: Values depict mean ± s.e.m. minimum independent pooled experiments. ns, significant, *P ≤ 0.05, **P 0.01, ***P 0.0001 ****P < 0.0001, unpaired, two-tailed test. data available online figure. 2 [embj2020107336-sup-0011-SDataFig2.xlsx] 3. A–E. Cartoon (top) illustrates larvae, disc (EAD, circle: green black), illustrated square (wild-type light brown atg13-mutant orange) (left) structure cephalic complex attached mouth hook. Larva (image larva microscope), (green highlights clones), (Phalloidin stains actin Hoechst blue nucleus), (Lipid red droplets, nucleus) top bottom. (A) 6. (B) (C) 8. (D) scrib−/−, atg13−/−//atg13−/− (E) ey3.5-atg13; complemented eye-specific transgenic atg13 expression, rescuing area space occupied lobes segments 4 (shown yellow dashed 2A), 45), 30), 30) 40), 25) 28). 7), 11) 13), 9) 9). Ventral Longitudinal (VL4) 2A) segment day8 11), 12) 14). I. atg13−/−//atg13−/−at 8) 7). J. measured mean±s.e.m. muscles 100 μm [embj2020107336-sup-0012-SDataFig3.xlsx] 4. Autophagy-driven releases metabolites circulation Changes groups (carbohydrates, acids, fatty acids) storage hemolymph progressing wasting, shown log2 LC-MS calculated per relative Volcano plot showing autophagy-dependent amounts X-axis shows fold change −/−scrib−/− // −/− vs. y-axis −log10 P-value, t-test. Metabolite names (FC) >± and/or –log10(P) Green points indicate 1, indicates −log10(P) 2, above thresholds. 113 reliably detected metabolites, those significant differences any comparisons shown. Color (fold change) difference numbers P-value comparison. statistical test define significance FDR-adjusted t-test 0.05. 5. Host-derived nutrients contribute amount glycogen biochemical normalized number B–E. ctrl, (white) illustrating (green) incorporation molecules (in blue). Sources incorporated differentiated changing isotopic content 25 h before measuring ratio tumor. A similar, experiment reported al (2021). 5 [embj2020107336-sup-0013-SDataFig5.xlsx] Click here expand EV1. A–C. D–F. (gray oval), mutant red) (no tumors), nuclei), nuclei) Wild type (w1118) control, spok-Gal4,UAS-Dcr2.D;UAS-rpr linger lack expressing (F) ecd1ts (ecdysoneless), lingers deficiency. muscle,