Different types of cartilage neotissue fabricated from collagen hydrogels and mesenchymal stromal cells viaSOX9,TGFB1orBMP2gene transfer
PLOS ONE
Authors: Weissenberger, Manuel; Weissenberger, Manuela H.; Wagenbrenner, Mike; Heinz, Tizian; Reboredo, Jenny; Holzapfel, Boris M.; Rudert, Maximilian; Groll, Juergen; Evans, Christopher H.; Steinert, Andre F.
Abstract
Objective As native cartilage consists of different phenotypical zones, this study aims to fabricate different types of neocartilage constructs from collagen hydrogels and human mesenchymal stromal cells (MSCs) genetically modified to express different chondrogenic factors. Design Human MSCs derived from bone-marrow of osteoarthritis (OA) hips were genetically modified using adenoviral vectors encoding sex-determining region Y-type high-mobility-group-box (SOX)9,transforming growth factor beta (TGFB) 1or bone morphogenetic protein (BMP) 2cDNA, placed in type I collagen hydrogels and maintained in serum-free chondrogenic media for three weeks. Control constructs contained unmodified MSCs or MSCs expressing GFP. The respective constructs were analyzed histologically, immunohistochemically, biochemically, and by qRT-PCR for chondrogenesis and hypertrophy. Results Chondrogenesis in MSCs was consistently and strongly induced in collagen I hydrogels by the transgenesSOX9,TGFB1andBMP2as evidenced by positive staining for proteoglycans, chondroitin-4-sulfate (CS4) and collagen (COL) type II, increased levels of glycosaminoglycan (GAG) synthesis, and expression of mRNAs associated with chondrogenesis. The control groups were entirely non-chondrogenic. The levels of hypertrophy, as judged by expression of alkaline phosphatase (ALP) and COL X on both the protein and mRNA levels revealed different stages of hypertrophy within the chondrogenic groups (BMP2>TGFB1>SOX9). Conclusions Different types of neocartilage with varying levels of hypertrophy could be generated from human MSCs in collagen hydrogels by transfer of genes encoding the chondrogenic factorsSOX9,TGFB1andBMP2. This technology may be harnessed for regeneration of specific zones of native cartilage upon damage.
HPV33+HNSCC is associated with poor prognosis and has unique genomic and immunologic landscapes
ORAL ONCOLOGY
Authors: Chatfield-Reed, Kate; Gui, Shanying; O'Neill, Wendi Q.; Teknos, Theodoros N.; Pan, Quintin
Abstract
Objective: To determine the influence of high-risk HPV genotype on outcomes in HNSCC patients. Materials and Methods: This is a retrospective analysis of The Cancer Genome Atlas HNSCC cohort. Results: Using multivariate Cox regression analysis, we revealed that HPV33 + HNSCC patients have inferior overall survival compared to HPV16 + HNSCC patients independent of anatomical site (HR 3.59, 95% CI 1.58-8.12; p = 0.002). A host anti-viral immune response, apolipoprotein B mRNA editing enzyme, and catalytic polypeptide-like mutational signature, was under represented and, aneuploidy and 3p loss were more frequent in HPV33 + tumors. A deconvolution RNA-Seq algorithm to infer immune cell fractions revealed that CD8 + cytotoxic T-cell infiltration was reduced in HPV33+ compared to HPV16 + tumors (1.3% vs. 2.7%, p = 0.007). TGFB1, a negative modulator of T-cell infiltration and function, showed expression and pathway enrichment in HPV33 + tumors. Conclusions: Our work reveals that HPV genotype, in particular HPV33, has a powerful impact on HNSCC patient survival. We argue that p16 immunohistochemistry as a surrogate biomarker for HPV + status will lead to sub-optimal risk stratification and advocate HPV genotype testing as standard of care.
COA
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