PBRM-1/PBAF-regulated genes in a multipotent progenitor in Caenorhabditis elegans.

The Caenorhabditis elegans somatic gonadal precursors (SGPs) are multipotent progenitors that generate all somatic cells of the adult reproductive system. The 2 SGPs originate in the mesodermal layer and are born through a division that produces one SGP and one head mesodermal cell (hmc). One hmc terminally differentiates, and the other dies by programmed cell death. The polybromo-associated BAF (PBAF) chromatin remodeling complex promotes the multipotent SGP fate. The complete loss of PBAF causes lethality, so we used a combination of Cre/lox recombination and GFP nanobody-directed protein degradation to eliminate PBRM-1, the signature subunit of the PBAF complex, from 83 mesodermal cells, including SGPs, body muscles, and the hmc. We used RNA sequencing to identify genes acting downstream of PBAF in these cells and identified 1,955 transcripts that were significantly differentially expressed between pbrm-1(-) and pbrm-1(+) in the mesoderm of L1 larvae. We found that genes involved in muscle cell function were overrepresented; most of these genes had lower expression in the absence of PBRM-1, suggesting that PBAF promotes muscle differentiation. Among the differentially expressed genes were 125 that are normally expressed at higher levels in SGP vs hmc and positively regulated by pbrm-1 and 53 that are normally expressed at higher levels in hmc vs SGP and are negatively regulated by pbrm-1; these are candidate regulators of the SGP/hmc fate decision. We validated one candidate gene using a fluorescent reporter; the hsp-12.3 reporter was derepressed in SGPs in pbrm-1 mutants, suggesting that hsp-12.3 expression is normally repressed by pbrm-1 in SGPs.

PUS7 deficiency in human patients causes profound neurodevelopmental phenotype by dysregulating protein translation.

Protein translation is a highly regulated process involving the interaction of numerous genes on every component of the protein translation machinery. Upregulated protein translation is a hallmark of cancer and is implicated in autism spectrum disorder, but the risks of developing each disease do not appear to be correlated with one another. In this study we identified two siblings from the NIH Undiagnosed Diseases Program with loss of function variants in PUS7, a gene previously implicated in the regulation of total protein translation. These patients exhibited a neurodevelopmental phenotype including autism spectrum disorder in the proband. Both patients also had features of Lesch-Nyhan syndrome, including hyperuricemia and self-injurious behavior, but without pathogenic variants in HPRT1. Patient fibroblasts demonstrated upregulation of protein synthesis, including elevated MYC protein, but did not exhibit increased rates of cell proliferation. Interestingly, the dysregulation of protein translation also resulted in mildly decreased levels of HPRT1 protein suggesting an association between dysregulated protein translation and the LNS-like phenotypic findings. These findings strengthen the correlation between neurodevelopmental disease, particularly autism spectrum disorders, and the rate of protein translation.

Primary lymphoma of the breast: A report of two cases.

Primary breast lymphoma (PBL) should be distinguished from secondary breast lymphoma arising in the setting of lymphoma elsewhere in the body. Multimodality imaging is key to diagnosing PBL, and imaging manifestations thereof may indicate PBL and alter the treatment course. Treatment options including chemotherapy, radiation therapy, and/or surgery depend on histology. We report two cases of PBL, illustrating the transformative impact that multimodality imaging may have on clinical management.

Acromioclavicular joint injuries and reconstructions: a review of expected imaging findings and potential complications.

Shoulder injuries, including acromioclavicular (AC) joint separations, remain a common reason for presentation to the emergency room. Although the diagnosis can be made apparent through proper history and physical examination by the emergency medicine physician, ascertaining the degree of injury can be difficult on the basis of clinical evaluation alone. While there is consensus in the literature that low-grade AC joint injuries can be treated with conservative management, high-grade injuries will generally require surgical intervention. Furthermore, the treatment of grade 3 injuries remains controversial, making it incumbent upon the radiologist to become comfortable with distinguishing this diagnosis from lower or higher grade injuries. Imaging of AC joint injuries after clinical evaluation is generally initiated in the emergency room setting with plain film radiography; however, on occasion, an alternative modality may be presented to the emergency room radiologist for interpretation. As such, it remains important to be familiar with the appearance of AC joint separations on a variety of modalities. Another possible patient presentation in both the emergent and nonemergent setting includes new onset of pain or instability in the postsurgical shoulder. In this scenario, the onus is often placed on the radiologist to determine whether the pain or instability represents the sequelae of reinjury versus a complication of surgery. The purpose of this review is to present an anatomically based discussion of imaging findings associated with AC joint separations as seen on multiple modalities, as well as to describe and elucidate a variety of potential complications which may present to the emergency room radiologist.

Cancer vaccine development: protein transfer of membrane-anchored cytokines and immunostimulatory molecules.

Many tumor cells escape host-immune recognition by the downregulation or lack of immunostimulatory molecules. Expression of immunostimulatory molecules on tumor cells by gene transfer can be used to induce an antitumor immune response. However, we have previously shown that protein transfer of glycosyl-phosphatidylinositol (GPI)-linked costimulatory molecules is a successful alternative to traditional gene transfer in preparing such a tumor vaccine. Vaccination with membranes modified by protein transfer to express GPI-linked B7.1 (CD80), a costimulatory adhesion molecule, induces protective immunity in mice and allogeneic antitumor T-cell proliferation in humans in vitro. Our goal is to develop an optimal tumor vaccine using tumor membranes modified by protein transfer to target and stimulate antigen-presenting cells (APCs) and T cells. We have investigated the efficacy of expressing GPI-anchored cytokine molecules on the surface of tumor cells. Expression of interleukin-12 (IL-12) on tumor-cell membranes in a GPI-anchored form induces a strong antitumor immune response that is comparable to the effects of secretory IL-12. Because many cytokines act synergistically, we are testing the membrane expression and immunostimulatory effects of cytokines individually as well as in combination to determine potential complementary effects of coexpression on the antitumor immune response. Ultimately, the protein-transfer vaccination may be used in humans alone or in multimodal combination therapies to induce tumor regression and to serve as a protective measure to prevent postsurgical secondary metastases.