https://openworks.mdanderson.org/sumexp21/1187/thumbnail.jp

Sadullozoda, Mumina

Savage, Hannah

Schadler, Keri L

English

OpenWorks @ MD Anderson

MethodsYUMMER melanoma cell line was injected into fourteen C571Bl/6 male mice subcutaneously. When tumors became palpable, mice were exercised 45 min/day, 5 days/week, for 14 days at 12 meters/minute using a forced treadmill model to achieve moderate aerobic exercise training. Prior to mouse euthanasia, pimonidazole HCl was injected intravenously in tumor bearing mice and tissue was frozen for analysis. Hypoxia and metabolism related proteins and genes were measured by western blot and qPCR, respectively. Differentially expressed metabolites were mined in a public dataset (Rundqvist et al. 2020. eLife) using the limmapackage on R.ConclusionsOur original hypothesis was not supported as we saw an increase in several metabolism related genes. These results suggest that aerobic exercise does modulate metabolism, however, it is unclear why exercise may increase metabolism in tumor tissue and how this impacts tumor hypoxia. Additional studies like seahorse assay to more directly determine metabolic flux and oxygen consumption are necessary to further elucidate the role of exercise in tumor metabolism.References Runqvist, H. et al. ELife (2020)Ryan, D.G. et al. Nat Metab 1, 16–33 (2019)Schadler, Keri L et al. Oncotarget vol. 7,40 (2016): 65429-65440.Title of the Poster Presentation Goes HereAuthors of the Poster Presentation Goes HereInstitutional and/or Graduate School of Biomedical Sciences Affiliation Goes Here The Effect of Aerobic Exercise on Tumor Hypoxia and Metabolism in a Murine Melanoma Model*Mumi a Sadullozoda, ٭Hannah Savage, ٭Keri L. Schadler*CUNY Brooklyn College, ٭Department of Pediatrics Research, MD Anderson Cancer CenterIntroduction Solid tumor vasculature is dysfunctional containing structural abnormalities, heterogeneous perfusion patterns, and hyper-permeability. These abnormalities make it a challenge to deliver systemic therapies effectively. In addition, poor vasculature function promotes a hypoxic tumor environment conducive to therapeutic resistance. One method to normalize tumor vasculature is aerobic exercise. Our lab has observed significant reductions in hypoxia after exercise but no change in vessel perfusion in a murine model of melanoma. This may be due to an alteration in tumor metabolic flux or oxygen consumption after exercise. Thus, this project aims to study the effect of exercise on tumor metabolism. We hypothesized that exercise reduces expression of genes associated with metabolism in tumors, contributing to decreased hypoxia within the tumor microenvironment.SedentaryExercise010203040YUMMERHypoxiap=0.05Hypoxyprobe: DAPI area *SedentaryExercise050100YUMMERVessel perfusionp=0.78Lectin: CD31 areaFig. 3 YUMMER melanoma subcutaneous tumor. Tumor was stained with CD31 (A) to measure vessel perfusion. Mice were injected with pimonidazole prior to sacrifice (B) to measure hypoxia. Fig. 1  Experimental design. Mice we randomized few days after the injection and divided into control and experimental groups.  YUMMER melanoma cell line injected to each miceExercise Mice for 14 days, 45 min/day, 5 times/week at 12 m/min Control = SedentaryN = 14EXERCISESEDENTARYPimonidazoleDAPILectin CD31A. Vessel Perfusion B. Hypoxia A BA BA BFig. 4 YUMMER melanoma tumor bearing mice were exercised or sedentary. Tumor samples were collected upon euthanasia. Tumors were lysed and western blotting was performed for HIF1αprotein normalized with β-actin. HIF1α gene was quantified using RT-qPCR. HIF1αβ-actinYUMMERControl ExerciseSedentaryExercise0.00.51.01.5HIFα Proteinp=0.87Band density (Normalized to B-actinl)AcknowledgementsDr. Jonghae Lee for western blot dataCPRIT-CURE Program for the research opportunity Results• Exercise reduced tumor hypoxia, as indicated by pimonidazole staining. However, no changes in HIF1α mRNA or protein expression were detected in tumor homogenates. Decreasing trends were observed in mRNA levels of HIF1αtarget genes, VEGFa and Glut1.• Genes associated with Krebs cycle function showed increasing trend in tumors from exercised mice (Idh2, p=0.09; and Fh1, p=0.16).• No change in expression of ATP synthase related genes were observed (ATP5b, p=0.37; ATPaf2, p=0.27). However, expression of cytochrome c oxidase increased after exercise (Cox4i1, p=0.05).• Consistent with our findings in tumor homogenates, Krebs cycle metabolites including citrate, α-ketoglutarate, fumarate, and malate increased in mouse plasma samples after exercise.Fig. 5 qPCR data shows decreasing trend, but no significant changes in HIF1α target genes after exercise. Aerobic exercise decreased hypoxia but did not alter vessel perfusionHIF1α protein and mRNA changes after exercise were not detectedSedentaryExercise0.00.51.01.52.0HIFα mRNAp=0.27mRNA Level(Normalized with GAPDH)HIF1α target genes demonstrated decreasing trends after exerciseSedentaryExercise0123VEGFamRNA level(normalized to TBP)p=0.09SedentaryExercise0.00.51.01.52.02.5Glut1mRNA level(normalized to TBP)p=0.13Fig. 2  Krebs cycle. Fumarate, malate, and α-ketoglutarate. were mined from a publicly available metabolomics dataset to identify differentially expressed metabolites in sedentary and exercised mouse plasma. SedentaryExercise0.00.51.01.52.0FumarasemRNA level(normalized to TBP)p=0.16SedentaryExercise0.00.51.01.52.0IsocitrateDehydrogenase 2mRNA level(normalized to TBP)p=0.09A BCOXPHOS GenesSedentaryExercise0123ATP5bmRNA level(normalized to TBP)p=0.37SedentaryExercise01234Cytochrome COxidasemRNA level(normalized to TBP)p=0.05Krebs cycle genesFig. 6 Fumarase and isocitrate dehydrogenase 2 are Krebs cycle enzymes. Increasing trend but no significant changes were observed in Krebs cycle gene expression after exercise when measured by RT-PCR  Fig. 7 Cytochrome c oxidase, ATPaf2, and ATP5b are three of oxidative phosphorylation pathway enzymes. mRNA levels of cytochrome C oxidase were significantly increased by exercise while ATPaf2 and ATP5b were not changed. SedentaryExercise0123ATPaf2mRNA level(normalized to TBP)p=0.27Created using Biorender.comFig. 8 Publicly available metabolomics dataset was mined to identify differentially expressed metabolites in sedentary and exercised mouse plasma. For differential expression analysis, exercise mouse plasma metabolites were compared to sedentary mouse plasma metabolites.Krebs cycle metabolites fumarate, malate, α-ketoglutarate, aconitate, citrate, and isocitrate showed significant increase after exercise(Exercise/Sedentary)shutters tock.com

The Effect of Aerobic Exercise on Tumor Hypoxia and Metabolism in a Murine Melanoma Model

https://openworks.mdanderson.org/cgi/viewcontent.cgi?article=1187&context=sumexp21

The Effect of Aerobic Exercise on Tumor Hypoxia and Metabolism in a Murine Melanoma Model

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