Research

Dr. Makowski’s expertise lies in immunometabolism, primarily bioenergetics and metabolic reprogramming of macrophages in the adipose and tumor microenvironment. The research focus of the Makowski laboratory is to identify targets in immune cells in complex diseases such as cancer, atherosclerosis, and diabetes. Using pre-clinical models and human subjects, we have identified metabolites, growth factors, and kinases that are regulated by obesity in adipose depots, including the mammary gland. We have demonstrated that gain and loss of function of fatty acid transporter FATP1 and glucose transporter GLUT1 alter macrophage function in vitro, and affect macrophage biology in obesity and atherosclerosis models. We have particular expertise in the normal mammary microenvironment and demonstrated obesity-associated alterations to the normal, pre-neoplastic, and tumor microenvironment influencing breast cancer risk, burden, and progression. We demonstrated for the first time obesity-dependent increases in the HGF/cMet oncogenic pathway that were reversible by weight loss, which reduced tumor progression. Using pharmacologic inhibition of cMet, we also reduced triple negative tumor burden. Our current work focuses on metabolic reprogramming of macrophages and investigation of other immune cells in obesity-induced breast cancer progression. She has published over 50 manuscripts and invited reviews in journals such as Molecular Metabolism, Breast Cancer Research Treatment, JBC, Obesity (cover), Cancer Cell, Nature Communications, and eLife. I have served on planning committees at AACR (obesity and cancer ‘17), South East Lipid Research Conference Organizing committee annually, and ASBMB ‘18. Graduate students (4) and fellows (3) have received funding from NIH (NIH NRSA F32, NIH Integrative Vascular Biology T32 slot), industry (Sanofi-Aventis), and/or internal merit-based (University of North Carolina Chancellor’s Fellowship, Royster Society of Fellows). Undergraduates trained have gone to Harvard Med, Wash U Med, UNC Med, UNC Dental, and Harvard Chan School of Public Health. Finally, Dr. Makowski has served on several study sections including Tumor Microenvironment (TME) and a special emphasis panel for NCI’s Provocative Questions as well other foundations including American Heart Association and Mary Kay Foundation. In 2017, Dr. Makowski was recruited to University of Tennessee Health Science Center to continue her work on immunometabolism in the normal and tumor microenvironment.

 

Contribution to Science # Denotes students and postdoctoral fellows mentored by Makowski.

1) Established the role of obesity and weight loss in basal-like breast cancer using translational multi-disciplinary approaches.

Macrophage-cancer-image2

Obesity leads to many types of cancer, but mechanisms are unclear. Central findings: 1) Using a unique murine model that resembles human basal-like breast cancer (an aggressive triple negative breast cancer), we demonstrated that high fat diet-induced obesity drove aggressive tumor behavior – e.g., early tumor onset (latency) and tumor progression – potentially through growth factors and inflammatory cytokines regulated by adiposity (Sundaram BCRT ’13). 2) We identified the HGF/cMet pathway as an obesity-driven oncogenic target, which had not been previously identified in any cancer type as obesity sensitive. We published the first evidence for obesity-induced HGF and activation of its receptor cMet in the etiology of basal-like breast cancer, which could be reversed with weight loss (Sundaram Frontiers in Oncology ’14, Qin CCI ’16). 3) Obesity-driven activation of several kinases in pre-neoplastic lesions of the mammary gland were further detected using kinomics that could be reversed by weight loss (Qin CCI ’16). 4) We also demonstrated that obesity drives ovarian cancer progression (Makowski Gyn Oncol ’14) in a novel murine model and that BMI is associated with endometrial cancer in TCGA samples (Roque Gyn Oncol ’16). 5) We further provided evidence of the importance of stromal-epithelial interactions in triple negative breast cancer promotion using murine and human samples. Normal breast of lean and obese women is enriched for macrophages (Sun BCRT ’12). Influence/application of findings: These findings are seminal in the understanding that the obese microenvironment shapes cancer risk and progression through changes in stromal cells (fibroblasts, macrophages, mast cells), kinases, and inflammatory mediators. Based on these findings, I was awarded two grants including a highly competitive NCI Provocative Question R21 and Mary Kay Foundation grant to test the role of weight loss and pharmacologic inhibition of cMet in reducing BBC through reversal of obesity-associated factors. We further established lack of knowledge of breast cancer subtypes and risk through community interactions and focus groups and created outreach and education materials including a website/app through the UNC Breast Cancer (http://mybcrisk.org/) and the Environment Research Program (BCERP, http://sph.unc.edu/bcerp/about-unc-bcerp/).

  1. #Qin Y, #Sundaram S, #Essaid L, Miller SM, Darr DB, Galanko JA, Montgomery SA, Major B, Johnson GL, Troester MA, Makowski L. Weight loss reduces basal-like breast cancer through kinome reprogramming. 16:26 Cancer Cell International. 1 April 2016. PMID: 2704215. PMCID: PMC4818517
  2. #Cozzo AJ*, #Sundaram S*, #Ottavia Zattra O, #Qin Y, Freemerman AJ, #Essaid L, Darr DB, Montgomery SA, McNaughton KK, Ezzell JA, Galanko JA, Troester MA, Makowski L. cMET inhibitor crizotinib impairs angiogenesis and reduces tumor burden in the C3(1)-Tag model of basal-like breast cancer. Springer Plus Breast Cancer Collection Springerplus. 2016 Mar 19;5:348. PMID: 27057482 PMCID: PMC4799044 *denotes co-first authorship
  3. #Sundaram S, #Le TL, #Essaid L, Freemerman AJ, #Huang MJ, Galanko JA, McNaughton KK, Bendt KM, Darr DB, Troester MA, Makowski L. Weight loss prevents obesity-associated basal-like breast cancer progression: Role of hepatocyte growth factor. Frontiers in Oncology 2014 Jul 8;4:175. PMID: 25072025. PMC4085881
  4. #Sundaram S, Freemerman AJ, McNaughton KK, Galanko JA, Bendt KM, Darr DB, Perou CM, Troester MA, Makowski, L. Role of HGF in obesity-associated tumorigenesis: C3(1)-Tag mice as a model for human basal-like breast cancer. Breast Cancer Res Treat. 2013 Dec;142(3):489-503. doi: 10.1007/s10549-013-2741-5. Epub 2013 Nov 12. PMID:24218051. PMC3904507.

 

2) Demonstrated that metabolic reprogramming of substrate metabolism, including glucose transport, glycolysis, and lipid trafficking direct macrophage biology in obesity and atherosclerosis.

23-b-and-f-RGB-A9-GLUT1-overexp-1024x780

Macrophages (MΦs) infiltrate adipose tissue in obesity; however, many questions remain about how MΦ phenotypes are regulated within the adipose or vessel microenvironment. We hypothesized that reducing utilization of certain substrates by deleting transporters would alter immune function – in this way we could metabolically reprogram macrophages to alter the course of disease. Central findings: Our findings demonstrated that the availability of fuel substrate is indeed a modifier of macrophage plasticity through metabolic reprogramming. We reported that fatty acid transport protein FATP1 regulated macrophage lipid metabolism and glucose tolerance (Johnson, Molecular Metabolism ‘16). We showed that GLUT1 expression drove glucose uptake, reactive oxygen species generation, and inflammation in macrophages (Freemerman J Biol Chem ‘14- currently 166 citations). Using a novel model created in our lab, we discovered defects in macrophage infiltration and pro-inflammatory activation in the absence of macrophage GLUT1 when mice were made obese (Freemerman, in revision). Furthermore, with American Heart Association funding, we showed that 1) GLUT1 is central for the phagocytic capacity of macrophages and may contribute to necrosis in an atherosclerotic lesion (Zhao, in revision); and 2) FATP1 regulates the size and severity of atherosclerotic lesions (Zhao, Atherosclerosis ‘17). Influence/application of findings: These studies defined metabolic mechanisms in MΦs leading to inflammation, atherosclerosis, and glucose intolerance.

  1. #Johnson AR*, #Qin YY*, #Cozzo A, Freemerman AJ, #Huang MJ, #Zhao L, #Sampey BP, #Milner JJ, Beck MA, Edin ML, Zeldin D, Galanko JA, Lee DP, Fueger PT, Damania B, Bivins B, Stahl A, Wu Y, Mohlke K, Makowski L. Macrophage Fatty Acid Transporter 1 (FATP1) Drives Alternative Macrophage Polarization and Limits Obesity-Induced Inflammation. *denotes co-first authorship. Molecular Metabolism, 5 (2016) 506-526. PMID: 27408776.
  2. Torres AM, Makowski L, Wellen KE. Metabolism fine-tunes macrophage activation. Elife. 2016 Feb 19;5. pii: e14354. doi: 10.7554/eLife.14354. PMID: 26894957. PMCID:PMC4769164
  3. Freemerman AJ, #Johnson AR, #Sacks GN, #Milner JJ, Kirk EL, Troester MA, Macintyre AN, Goraksha-Hicks P, Rathmell JC, Makowski L. Metabolic reprogramming of macrophages: Glucose Transporter (GLUT1)-mediated glucose metabolism drives a pro- inflammatory phenotype. J Biol Chem. 2014 Feb 3. PMID: 24492615. PMC3953299.
  4. #Zhao L, #Cozzo AJ, #Johnson AR, #Christensen T, Freemerman AJ, Bear JE, Rotty JD, Bennett BJ, Makowski L. Lack of myeloid Fatp1 increases atherosclerotic lesion size in Ldlr-/- Atherosclerosis. 2017 Nov;266:182-189. Epub 2017 Oct 7. PMID: 29035781.

 

3) Established the inflammatory potential of varied diets, exposures, and specific metabolites.

Sampey Makowski Cover Obesity 2011

Inflammation is linked to obesity, diabetes and some cancers, but the specific mechanisms remain unclear. We hypothesized that inflammatory and metabolic biomarkers of risk associated with inflammation, insulin resistance, and diabetes could identify pathways to target in control of obesity-induced pathology. Central findings: We defined for the first time metabolic biomarkers of obesity-induced inflammation and beta-oxidative metabolism in adipose tissue (Sampey Obesity ‘11 – has 321 citations). We presented novel findings on macrophage infiltration into BAT tissue with a dramatic lack of crown like structures detected but significant macrophage staining around vessels where fat droplets accumulated. The Obesity manuscript was featured on the cover of the journal Obesity. Subsequently, using high throughput metabolomic analysis, we identified novel lipid metabolites as biomarkers of insulin resistance and adipose inflammation. We then demonstrated in vitro that a specific metabolite induced the production of classical pro-inflammatory cytokines associated with diabetes using bone marrow derived macrophages (Sampey, PLoS One ’12- has 78 citations). The PLOS One study was featured on National Public Radio. Gene expression associated with the CAF diet showed oxidative stress (Johnson BBRC 2016). Influence/application of findings: Cafeteria Diet model provides a unique platform to further study the biochemical, genomic and physiological mechanisms of obesity and obesity-related disease states that are pandemic in Western civilization today.

  1. #JohnsonAR, WilkersonMD, #SampeyBP, TroesterMA, HayesDN, Makowski L. Cafeteria Diet Induced Obesity Results in Increased Oxidative Damage in White Adipose. 2016 Mar 28. PMID: 27033600. PMCID: PMC4862365
  2. #Sampey BP, Freemerman AJ, #Zhang J, Kuan PF, Galanko JA, O’Connell TM, Ilkayeva OR, Muehlbauer MJ, Stevens RD, Newgard CB, Brauer HA, Troester MA, Makowski L. Metabolomic Profiling Reveals         Mitochondrial-Derived Lipid Biomarkers that Drive Obesity-Associated Inflammation. PLoS One. 2012;7(6):e38812. Epub 2012 Jun 12. PMID: 22701716. PMC3373493.
  1. #Sampey BP, Vanhoose AM, Winfield H, Freemerman AJ, Muehlbauer M, Fueger PT, Newgard CB, and Makowski L. Cafeteria-Diet is a Robust Model of Human Metabolic Syndrome with Liver and Adipose Inflammation: Comparison to High Fat Diet. Obesity (Silver Spring). Jun;19(6):1109-17. Epub 2011 Feb 17. PMID: 21331068. 2011. PMC3130193. Cover photo.