Funding Status:
Completed - Task completed and produced a deliverable
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Procurement Mechanism(s):
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Aims:
Aim 1. To determine the proteomic and phosphoproteomic changes which occur to rat neonatal
cardiomyocytes (RNNCM) upon exposure to microgravity. RNNCM will be placed in the
NASA bioreactor, also referred to as the Rotating Wall Vessel (RWV). RNNC were chosen for the first aim in this pilot as they are a readily attainable and hardy cell type which can withstand being cultured for weeks, for determining the minimum time needed for observable microgravity effects via proteomics analysis. We will evaluate proteomic changes at 1 week, 2 week, and 1 month of (+/-) exposure to microgravity.
Aim 2. To determine the proteomic/phosphoproteomic changes in human heart isolated trabeculae (iHT) following exposure to microgravity. Human hearts will be procured according to IRB approved protocols. iHT are thin multi-cellular tissues located on the endocardial surface of the human heart. They offer a unique advantage in that they are multicellular, contain extracellular matrix, and are thin enough to receive adequate oxygen and nutrients while contracting and developing force. We will assess human iHT proteomic differences in samples exposed to microgravity (using the optimal time point for microgravity effects obtained from RNNC pilot). Total # of samples=18 (9 samples from two individuals at 1 time point, +/-microgravity
exposure, ischemic injury treated control). In addition, the proteomic profile of the “secretome” (i.e. media in which the iHT are cultured) will be evaluated for each sample.
Together Aims 1 and 2 in the new proposal will provide evidence that the human heart tissue can be exposed to microgravity, provide dosage and timing of exposure for maximal proteomic changes, and will determine survival of the trabeculae in the apparatus.