The overall goal of this work is to apply in vivo and in vitro models to study the basement membrane protein laminin α4 (lama4) in adipocyte... Show moreThe overall goal of this work is to apply in vivo and in vitro models to study the basement membrane protein laminin α4 (lama4) in adipocyte function. Adipose tissue accumulation, lipogenesis, and structure were examined in mice with a null mutation of the lama4 gene (Lama4−/−) and compared to wild-type animals (Lama4+/+). The Lama4−/−mice phenotype was investigated to evaluate if the differences were due specifically to the adipose tissue function. Physical activity and food intake does not differ between Lama4+/+ and Lama4−/− mice. However, Lama4−/− mice have a significantly increased metabolic rate at 25°C and 16°C compared to Lama4+/+ mice. In contrast, in thermoneutral conditions at 30°C both Lama4+/+ and Lama4−/− mice exhibit equivalent metabolic rates. Interestingly, when room temperature housed mice fat pads were evaluated with immunohistochemistry, Lama4−/− mice exhibit significantly increased UCP-1 expression in subcutaneous adipose. These results suggest that beiging, white to brown adipocytes, in subcutaneous adipose tissue in Lama4−/− mice may lead to decreased adipose tissue accumulation and improved metabolic function. While animal models indicate the absence of lama4 results in more beiging in subcutaneous adipose tissue, an in vitro tissue engineered model was developed to study the adipocyte function in a controlled microenvironment. Primary cell spheroids developed from Lama4+/+ and Lama4−/− were incorporated into synthetic poly(ethylene glycol) (PEG) hydrogels within a range of stiffnesses. When the cells were given the same chemical cues their functions differed depending upon microenvironment stiffness. Beige function in adipocyte cells in 3D can be influenced by matrix stiffness. In conclusion, the laminin alpha 4 basement membrane protein absence in adipose tissue results in adipocyte functional changes in vivo. The Lama4−/− mice have resistance to diet induced weight gain and increased metabolic rate at room temperature and when cold challenged. The Lama4−/− mice had increased beiging in the subcutaneous depot. Additionally, a tissue engineered model was developed to further study cell-ECM interactions first identified in an animal model. These in vivo findings and the engineered model of adipose tissue have great potential for studying obesity and other adipose related diseases. Future work will require continued interdisciplinary collaboration towards the successful identification and screening of novel therapeutics using engineered tissue models. Ph.D. in Biomedical Engineering, July 2015 Show less
