Cell & Molecular Biology
Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/30806
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Browsing Cell & Molecular Biology by Author "Rinderle, Caroline"
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Item Characterization of Estrogen Receptors (ERs) and ER-metabolizing enzymes in Lipedema and Non-Lipedema Adipose Stem Cells (ASCs) and differentiated adipocytes(2022) Walczak, Samantha; Al-Ghadban, Sara; Rinderle, Caroline; Bunnell, BruceIntroduction: Lipedema is a chronic, idiopathic painful disease characterized by an excess of adipose tissue in the lower extremities, commonly misdiagnosed as obesity, lymphedema, or chronic venous insufficiency. As the severity of lipedema worsens patients have reduced mobility, easy bruising, and fatigue and it is thought to resist lifestyle modifications. While treatments such as liposuction can help ease these symptoms, it is not curative, and the underlying etiology is unknown. Hypothesis: As the development of lipedema often begins or worsens during periods of hormonal change such as puberty, pregnancy, or menopause, we hypothesize that alterations in estrogen drive lipedema pathogenesis. Aim: The aim of this study is to characterize the gene expression of estrogen receptors (ER-α and ER-β), G-protein coupled estrogen receptor (GPER), and ER metabolizing enzymes: Hydroxysteroid 17-beta dehydrogenase (HSD17B1, B7, B12), Hormone-sensitive Lipase (LIPE) and Steroid Sulfatase (STS) in ASCs and differentiated adipocytes in BMI and age-matched non-lipedema and lipedema patients. Methods: Cell culture and Oil Red O stain, RNA extraction and RT-PCR assays were used to assess the expression of ERs and the estrogen metabolizing enzymes in ASCs and differentiated adipocytes. Results: ER-α, ER-β, and GPER gene expression were increased in Lipedema ASCs cultured in hormone-depleted media, as well as in differentiated adipocytes compared to non-lipedema corresponding cells. LIPE, STS, HSD17B17, and HSD17B12 gene expression were also increased in Lipedema differentiated adipocytes compared to non-lipedema differentiated adipocytes. In addition, the gene expression of HSD17B1 was increased in Lipedema ASCs cultured in hormone-depleted media compared to non-lipedema ASCs. Conclusion: These results indicate that expression of ERs and estrogen metabolizing enzymes are altered by Lipedema and suggest that estrogen may play a role in adipose tissue dysregulation in lipedema. Exploring this possible etiology further could contribute to the expansion of treatment options and management available to lipedema patients.Item Novel Kinase Inhibitors Library Screen Differentially Impacts Adipose Stem Cells (ASCs) from Lean and Obese Donors(2022) Rinderle, CarolinePurpose: Breast cancer is the second leading cause of death among women in the United States. Obesity increases the risk of developing breast cancer and ultimately leads to poorer outcomes. Obesity is defined as the excess accumulation of adipose tissue, evidenced by a BMI of 30 kg/m² or greater. Adipose tissue consists primarily of adipocytes, but the stromal vascular fraction (SVF) consists of numerous other cell types as well, including adipose-derived stem cells (ASCs). ASCs are self-renewing, multipotent, mesenchymal stem cells that have been intensely studied for their role in regenerative medicine. Our lab has shown that ASCs extracted from obese patients are recruited to breast tumors more than ASCs extracted from lean patients. This made breast cancer outcomes worse in our xenograft models, indicating a connection between ASCs in obesity and breast cancer. These changes in cancer behavior may be due to the activity of important protein kinases. Kinases are essential to cellular function, activating necessary proteins to propagate signal cascades, without which survival would be impossible. Little is known about the 538 kinases encoded in the human genome, and therefore, they need to be researched more thoroughly. If the obese ASC-breast tumor crosstalk can be interrupted via prevention of specific kinase activity, then poor breast cancer outcomes may be prevented and novel therapeutics can be uncovered. Methods: Pools of ASCs from lean and obese donors were treated 100nM of KCGS Drug Library kinase inhibitors obtained from Dr. David Drewry at the SGC at UNC Chapel Hill. After 72 hours, cells were stained with crystal violet and imaged for cellular viability and morphologic changes. Results: Fifteen kinase inhibitor drugs affected ASCs from both lean and obese donors. Nine kinase inhibitor drugs affected ASCs from obese donors only. No kinase inhibitors affected ASCs from lean donors alone. Conclusions: Obesity changes the biology of ASCs in a way that can potentially be therapeutically targeted. In the future, testing the ability of ASCs to differentiate into mature adipocytes after treatment with kinase inhibitors will give greater insight into the role specific kinases play in the biology of an obese patient. We will also study the roles of the kinase inhibitors and kinases in the biology of ASCs more thoroughly.