Final publication is available at http://www.sciencedirect.com/science/article/pii/S0006291X1530276X / 京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19563号 / 医博第4070号 / 新制||医||1013(附属図書館) / 32599 / 京都大学大学院医学研究科医学専攻 / (主査)教授 河本 宏, 教授 藤田 潤, 教授 髙折 晃史 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Skeletal muscle associated disorders are correlated with significant morbidity, including frailty, fatigue, reduced mobility and poor resistance to treatments as well as mental health repercussions resulting from a loss of independence. Thus, conditions affecting skeletal muscle put considerable pressure on the health care system. In response to injury, skeletal muscle can regenerate and the molecular mechanisms underlying this unique process has been the subject of intense research with the goal of developing better treatment modalities for muscle-related diseases. Our laboratory has previously demonstrated that C/EBPβ is a negative regulator of postnatal myogenic differentiation. Expressed in muscle satellite cells (MuSCs), the primary source of regenerative potential in skeletal muscle, C/EBPβ inhibits entry into the myogenic differentiation program and is required for MuSC self-renewal after injury. Despite the important role of C/EBPβ in muscle homeostasis, little is known about the genes it regulates. To better understand how C/EBPβ regulates these processes, I used both a candidate-based approach to identify the inhibitor of DNA binding and differentiation protein ID3 as a C/EBPβ target gene that mediates inhibition of myogenic differentiation, and an unbiased approach using RNA-seq. I compared gene expression profiles from C2C12 myoblasts overexpressing C/EBPβ to control cells under growth and differentiation conditions. I observed that more than 20% of the molecular signature found in quiescent MuSCs is regulated by C/EBPβ. Caveolin- 1 was implicated as a direct target of C/EBPβ and part of the molecular mechanism by which C/EBPβ maintains MuSCs quiescence. Interestingly, the RNA-seq data identified numerous C/EBPβ-regulated secreted proteins including growth factors and cytokines. Co-culture experiments indicate that secreted proteins mediate the inhibition of cell differentiation and fusion, suggesting that C/EBPβ functions in an autocrine and paracrine fashion to influence activation of myoblasts in the absence of cell-to-cell contact. Given the role of C/EBPβ in regulating secretory proteins that inhibit myogenic differentiation, I examined the requirement of C/EBPβ in the expression of anti myogenic proteins secreted by cancer cells that affect MuSCs function and drive the development of cancer cachexia. Indeed, the expression of C/EBPβ in cancer cells was found to be required for the production of a cachexia-inducing secretome by tumours in vitro and in vivo. Furthermore, C/EBPβ was found to be sufficient to convert non-cachectic tumours into cachexia-inducing ones. In comparing differentially expressed C/EBPβ-regulated secreted protein transcripts from our RNA-seq data to that from 27 different types of human cancers revealed an ~18% similarity between C/EBPβ-regulated secreted proteins and those enriched in cachectic tumours including pancreatic, gastric and brain cancers. Three of these C/EBPβ-regulated secreted proteins (SERPINF1, TNFRSF11B and CD93) were tested further and found to be inducers of muscle atrophy. This work provides molecular insight into the role of C/EBPβ in the regulation of MuSC function and the regulation of cachexia-inducing factors by tumours, placing C/EBPβ as a novel therapeutic target for the treatment of cancer cachexia and other muscle-related diseases.