Katherine M Marsh1, David Schipper1, Alice S Ferng1, Kitsie Johnson1, Julia Fisher2, Shannon Knapp2, Destiny Dicken1, Zain Khalpey3,4,5. 1. Division of Cardiothoracic Surgery, Department of Surgery, University of Arizona College of Medicine, Tucson, AZ, USA. 2. Statistical Consulting Lab, BIO5 Institute, University of Arizona, Tucson, AZ, USA. 3. Division of Cardiothoracic Surgery, Department of Surgery, University of Arizona College of Medicine, Tucson, AZ, USA. zkhalpey@surgery.arizona.edu. 4. Department of Translational and Regenerative Medicine, University of Arizona College of Medicine, Tucson, AZ, USA. zkhalpey@surgery.arizona.edu. 5. , 1501 North Campbell Avenue, Room 4302, Tucson, AZ, 85724, USA. zkhalpey@surgery.arizona.edu.
Abstract
INTRODUCTION: Lymphangioleiomyomatosis (LAM) is a rare, progressive cystic lung disease that predominantly affects women of childbearing age. Exogenous rapamycin (sirolimus) has been shown to improve clinical outcomes and was recently approved to treat LAM, whereas estrogen (E2) is implicated in disease progression. No consistent metabolic model currently exists for LAM, therefore wild-type mouse embryonic fibroblasts (MEF +/+) and TSC2 knockout cells (MEF -/-) were used in this study as a model for LAM. METHODS: Oxygen consumption rates (OCR) and redox potential were measured to determine metabolic state across control cells, MEF +/+ and -/- cells treated with rapamycin (Rapa), and MEF +/+ and -/- cells treated with E2. An XF96 extracellular flux analyzer from Seahorse Bioscience® was used to measure OCR, and a RedoxSYS™ ORP was used to measure redox potential. RESULTS: OCR of MEF -/- cells treated with rapamycin (MEF -/- Rapa) versus MEF -/- control were significantly lower across all conditions. The static oxidation reduction potential of the MEF -/- Rapa group was also lower, approaching significance. The coupling efficiency and ratio of ATP-linked respiration to maximum respiration were statistically lower in MEF -/- Rapa compared to MEF +/+ Rapa. There were no significant metabolic findings across any of the MEF cells treated with E2. MEF -/- control cells versus MEF +/+ control cells were not found to significantly differ. CONCLUSION: MEF cells are thought to be a feasible metabolic model for LAM, which has implications for future pharmacologic and biologic testing.
INTRODUCTION:Lymphangioleiomyomatosis (LAM) is a rare, progressive cystic lung disease that predominantly affects women of childbearing age. Exogenous rapamycin (sirolimus) has been shown to improve clinical outcomes and was recently approved to treat LAM, whereas estrogen (E2) is implicated in disease progression. No consistent metabolic model currently exists for LAM, therefore wild-type mouse embryonic fibroblasts (MEF +/+) and TSC2 knockout cells (MEF -/-) were used in this study as a model for LAM. METHODS:Oxygen consumption rates (OCR) and redox potential were measured to determine metabolic state across control cells, MEF +/+ and -/- cells treated with rapamycin (Rapa), and MEF +/+ and -/- cells treated with E2. An XF96 extracellular flux analyzer from Seahorse Bioscience® was used to measure OCR, and a RedoxSYS™ ORP was used to measure redox potential. RESULTS: OCR of MEF -/- cells treated with rapamycin (MEF -/- Rapa) versus MEF -/- control were significantly lower across all conditions. The static oxidation reduction potential of the MEF -/- Rapa group was also lower, approaching significance. The coupling efficiency and ratio of ATP-linked respiration to maximum respiration were statistically lower in MEF -/- Rapa compared to MEF +/+ Rapa. There were no significant metabolic findings across any of the MEF cells treated with E2. MEF -/- control cells versus MEF +/+ control cells were not found to significantly differ. CONCLUSION:MEF cells are thought to be a feasible metabolic model for LAM, which has implications for future pharmacologic and biologic testing.
Authors: Jane J Yu; Victoria A Robb; Tasha A Morrison; Eric A Ariazi; Magdalena Karbowniczek; Aristotelis Astrinidis; Chunrong Wang; Lisa Hernandez-Cuebas; Laura F Seeholzer; Emmanuelle Nicolas; Harvey Hensley; V Craig Jordan; Cheryl L Walker; Elizabeth P Henske Journal: Proc Natl Acad Sci U S A Date: 2009-02-06 Impact factor: 11.205
Authors: Xiaoxiao Gu; Jane J Yu; Didem Ilter; Nickolas Blenis; Elizabeth Petri Henske; John Blenis Journal: Proc Natl Acad Sci U S A Date: 2013-08-27 Impact factor: 11.205
Authors: Chenggang Li; Po-Shun Lee; Yang Sun; Xiaoxiao Gu; Erik Zhang; Yanan Guo; Chin-Lee Wu; Neil Auricchio; Carmen Priolo; Jing Li; Alfredo Csibi; Andrey Parkhitko; Tasha Morrison; Anna Planaguma; Shamsah Kazani; Elliot Israel; Kai-Feng Xu; Elizabeth Petri Henske; John Blenis; Bruce D Levy; David Kwiatkowski; Jane J Yu Journal: J Exp Med Date: 2014-01-06 Impact factor: 14.307