Osteogenesis imperfecta (OI) is a heterogeneous group of genetic disorders that affect connective tissue integrity. The hallmark of OI is bone fragility, although other manifestations, which include osteoporosis, dentigenesis imperfecta, blue sclera, easy bruising, joint laxity and scoliosis, are also common among OI patients. The severity of OI ranges from prenatal death to mild osteopenia without limb deformity. Most forms of OI result from mutations in the genes that encode either the proa1or proa2 polypeptide chains that comprise type I collagen molecules, the major structural protein of bone. Treatment depends mainly on the severity of the disease with the primary goal to minimize fractures and maximize function. Current treatments include surgical intervention with intramedullarly stabilization and the use of prostheses. Pharmacological agents have also been attempted with limited success with the exception of recent use of bisphosphonates, which have been to shown to have some effect. Since OI is a genetic disease, these agents are not expected to alter the course of the collagen mutations. Cell and gene therapies as potential treatments for OI are therefore currently being actively investigated. The design of gene therapies for OI is however complicated by the genetic heterogeneity of the disease and by the factor that most of the OI mutations are dominant negative where the mutant allele product interferes with the function of the normal allele. The present review will discuss the molecular changes seen in OI, the current treatment options and the gene therapy approaches being investigated as potential future treatments for OI.

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Purpose The Serratia marcescens bacterium causes vision‐threatening keratitis, life‐threatening hospital acquired infections and contaminates contact lens cases. The goal of this study was to identify a hemolytic factor produced by keratitis isolates and laboratory strains of S. marcescens. Methods Hemolysis was measured using sheep and mouse erythrocytes. S. marcescens was mutated with a mariner transposon. Hemolysin defective mutants were isolated on blood agar plates. Transposon insertion sites were mapped by marker rescue and sequencing. Complementation analysis was performed with plasmids. Serratamolide was extracted with ethyl acetate and purified by preparative HPLC and verified as pure with high‐resolution mass spectroscopy (HR‐MS) and 1H NMR analysis. Cytotoxicity to human airway and ocular epithelial cells was performed using Alamar Blue viability dye. The presence of swrW in ocular isolates was determined using PCR. Results Mutation of the swrW gene conferred a hemolysis defect that was complemented by the wild‐type swrW gene on a plasmid. The SwrW protein catalyzes production of the cyclic lipopeptide serratamolide. Purified serratamolide was hemolytic to mammalian erythrocytes, and cytotoxic to human airway and ocular cells in vitro. The swrW gene was found in the majority of contact‐lens associated keratitis isolates. Conclusion Serratamolide is a novel hemolysin produced by S. marcescens ocular isolates and may contribute to the ability of contact lens associated bacteria to cause infections.

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BACKGROUND: Ovarian cancer (OC) is the deadliest gynecologic cancer. Despite randomized clinical trials showing improved survival for patients receiving intraperitoneal chemotherapy (IP), IP has not received a wide use outside of specialty hospitals. The aim of this study was to explore the impact of IP on OC patient survival and to evaluate whether treatment facility type impacted the outcomes. METHODS: Detailed demographic and clinical information on OC patients (N=2924) who underwent treatment in UPMC facilities between 2000-2016 was obtained from the UPMC Cancer Registry. Duplicate records, rare, grade 1, and non-epithelial tumors were excluded. Kaplan-Meier plots were constructed to compare 10-year survival rates based on the chemotherapy type (IP vs. no-IP), cancer grade, cancer stage, surgery type, neoadjuvant and treatment facility (specialized vs. community). Multivariable Gray's models and logistic regression model were fitted to evaluate the effect of different factors on survival and tumor recurrence respectively. Two tailed P-values <0.05 were considered significant. R software package was used to analyze the data. RESULTS: The final sample consisted from 1840 patients (250 IP and 1590 no-IP). IP chemotherapy was used only in 14% of OC patients and was associated with improved long-term survival. Similarly, cases reported by specialty treatment facilities (Magee and Passavant) had better survival compared to other hospitals. Multivariable Gray's model showed that IP and younger age were significantly associated with lower hazard of death, whereas higher cancer stage is significantly associated with higher hazard of death. Multivariable logistic regression showed that IP is not significantly associated with recurrence after adjusting for median income in the zip code of patient residence and cancer stage. CONCLUSIONS: These findings demonstrate enhanced long-term survival of patients treated with IP therapy at specialty centers. Increasing IP therapy use in clinical practice for OC patient treatment may be important strategy to improve OC outcomes. Citation Format: Adambekov S, Wang S, Taylor S, Buckanovich R, Coffman L, Edwards RP, Linkov F. INTRAPERITONEAL CHEMOTHERAPY USE AND OUTCOMES: EXPLORATION OF UPMC OVARIAN CANCER REGISTRY DATA [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr DP-010.

There is increasing interest in adeno-associated virus (AAV) vectors for a wide variety of gene therapy applications. AAV is a nonpathogenic human parvovirus that can mediate long-term transduction of a number of cell types without provoking a significant immune response. These properties make AAV especially attractive for use in gene therapy of rheumatoid arthritis (RA), a chronic inflammatory disease. To investigate the potential of AAV in gene therapy of arthritis, the ability of AAV to infect synovium in vitro and in vivo was tested. Three human RA synovial fibroblast cell lines and two murine (one DBA/1J and one DBA1J×C3H F1) synovial fibroblast cell lines were used to test AAV transduction in vitro. The cell lines (2 × 10 5 cells) were infected with 10 4 particles/cell of a murine IL-10-encoding vector (AAV-mIL-10) alone or with the addition of a low titer (100 particles/cell) of an E1-, E3-deleted recombinant adenovirus to provide E4orf6 activity to enhance second-strand synthesis. The supernatants were harvested from the wells at various time points and assayed for mIL-10 expression by ELISA. Both human synovial cell lines infected with AAV alone demonstrated low-level transgene expression throughout the course of the study. However, by day 10, all human cultures coinfected with adenovirus showed a 16-to 56-fold increase in mIL-10 compared to cultures infected with AAV-mIL10 alone. By day 30, a 31-to 135-fold increase was observed. No such increase was observed in any of the mouse cell lines. To determine the AAV transduction efficiency for synovium in vivo, human RA synovial tissues obtained from patients undergoing joint-replacement surgery were implanted subcutaneously on the backs of NOD.CB17-Prkdc SCID mice. After allowing a 2-week period for engraftment, tissues were injected with 3.4 × 10 11 particles of AAV-luciferase alone or in combination with 1.0 × 10 11 particles of adenovirus. Two weeks following AAV administration, the tissues were homogenized and assayed for expression of luciferase. Only the tissues coinfected with adenovirus had luciferase levels above background. A similar experiment with AAV-LacZ demonstrated X-gal staining only of synovial tissues coinfected with adenovirus. These findings demonstrate a preferential ability of AAV to transduce human, compared to mouse, synovial tissue and suggest that second strand synthesis may be a limiting factor in gene transduction. Further studies to elucidate the mechanisms limiting gene transduction in human synovium may allow optimization of this vector for the treatment of arthritis. Research of the last years has demonstrated clearly the role of rheumatoid arthritis synovial fibroblasts (RA-SF) in the destruction of articular cartilage. It has been understood that RA-SF not only exhibit features of activation and altered apoptosis, but following attachment to cartilage secrete large amounts of matrix degrading enzymes that mediate the destruction of extracellular matrix. Given recent advances in the field of gene transfer, we have been wor...