Britson, K. A. | Search bioRxiv

bioRxiv2024

Arc controls organ architecture through modulation of Crb and MyoII

Precise orchestration of morphogenetic processes is required to generate organs that are optimally situated within the organism, and that are of the right size and shape to fit and maximize functionality. Here, we describe the role of Arc, a large apical membrane-associated PDZ domain-containing protein, that works through the apical determinant Crumb (Crb) to limit MyoII activity during tissue invagination in the forming Drosophila salivary gland (SG). We show that loss of Arc, attenuation of Crb function, as well as increased activation of non-muscle Myosin II (MyoII) leads to the simultaneous internalization of more precursor cells than normal. Consequently, mature SGs are significantly shorter with more cells surrounding the lumen at all positions along the tube. Correspondingly, overexpression of Arc or SG-specific knockdown of MyoII leads to the formation of longer SGs with fewer cells surrounding the lumen. We show that both Arc PDZ domains are required for Arc function and that they have distinct activities. Finally, we show that Arc facilitates Crb plasma membrane (PM) localization and suggests a model wherein PM-associated Crb stabilizes cellular junctions countering the destabilizing effects of apical medial and junctional pools of activated MyoII, thus limiting the number of primordial cells internalizing at any given time.

developmental biology↗

bioRxiv2021

Loss of TDP-43 function and rimmed vacuoles persist after T cell depletion in a xenograft model of sporadic inclusion body myositis

Sporadic inclusion body myositis (IBM) is the most common acquired muscle disease in adults over age 50, yet it remains unclear whether the disease is primarily driven by T cell-mediated autoimmunity. IBM muscle biopsies exhibit nuclear clearance and cytoplasmic aggregation of TDP-43 in muscle cells, a pathologic finding observed initially in neurodegenerative disease, and nuclear loss of TDP-43 in neurons causes aberrant RNA splicing. Here, we show that loss of TDP-43 splicing repression, as determined by inclusion of cryptic exons, occurs in skeletal muscle of IBM patients. Out of 119 muscle biopsies tested, RT-PCR-mediated detection of cryptic exon expression is 84% sensitive and 99% specific for diagnosing IBM, indicating utility as a functional and diagnostic biomarker. To determine the role of T cells in pathogenesis, we generated a novel xenograft model by transplanting human IBM muscle into the hindlimb of immunodeficient mice. Xenografts from IBM patients display robust regeneration of human myofibers and recapitulate both inflammatory and degenerative features of the disease. Myofibers in IBM xenografts are invaded by human, oligoclonal CD8+ T cells and exhibit MHC-I upregulation, rimmed vacuoles, mitochondrial pathology, p62-positive inclusions, and nuclear clearance and cytoplasmic aggregation of TDP-43, resulting in expression of cryptic exons. Depletion of human T cells within IBM xenografts by treating mice intraperitoneally with anti-CD3 (OKT3) suppresses MHC-I upregulation, but rimmed vacuoles and loss of TDP-43 function persist. These data suggest that myofiber degeneration occurs independent of T cells, and muscle cell-intrinsic mechanisms, such as loss of TDP-43 splicing repression, drive IBM pathogenesis. One Sentence SummaryDepletion of T cells in a xenograft model of sporadic inclusion body myositis suppresses inflammation but not TDP-43 pathology or muscle degeneration.

pathology↗