Short hairpin (sh)RNAs delivered by recombinant adeno-associated viruses (rAAVs) are valuable tools to study gene function in vivo and a promising gene therapy platform. Our data show that incorporation of shRNA transgenes into rAAV constructs reduces vector yield and produces a population of truncated and defective genomes. We demonstrate that sequences with hairpins or hairpin-like structures drive the generation of truncated AAV genomes through a polymerase redirection mechanism during viral genome replication. Our findings reveal the importance of genomic secondary structure when optimizing viral vector designs. We also discovered that shDNAs could be adapted to act as surrogate mutant inverted terminal repeats (mTRs), sequences that were previously thought to be required for functional self-complementary AAV vectors. The use of shDNAs as artificial mTRs opens the door to engineering a new generation of AAV vectors with improved potency, genetic stability, and safety for both preclinical studies and human gene therapy.

Background/Aims: Hepatocellular carcinoma (HCC) is one of the most deadly diseases; metastasis and recurrence are the most important factors that affect the therapy of HCC chronically. Until now, the prognosis for the metastasis of HCC had not improved. Recently, several proteins that are related to metastasis and invasion of HCC were identified, but the effective markers still remain to be elucidated. Methods: In this study, comparative proteomics was used to study the differentially expressed proteins in two HCC cell lines MHCC97L and HCCLM9, which have low and high metastatic potentials, respectively. Results: Our findings indicated that filamin A (FLNA) and phosphoglycerate kinase 1 (PGK1) were two significantly differentially expressed proteins, with high expression in HCCLM9 cells, and may influence the metastasis of HCC cells. Conclusion: Taken together with the confirmation of expression on the mRNA level, we propose the use of FLNA and PGK1 as potential markers for the progression of HCC.

To assess the prognostic impact of pretreatment albumin-to-alkaline phosphatase ratio (AAPR) in patients with upper tract urothelial carcinoma (UTUC), the data of 692 patients, operated between 2003 and 2016 in our center, were retrospectively assessed. The threshold of AAPR was defined as 0.58 by using the receiver-operating curve analysis. Overall survival (OS), cancer-specific survival (CSS), and recurrence-free survival (RFS) were evaluated using the Kaplan-Meier method. And the univariate and multivariate Cox’s regression models were performed to identify independent prognostic predictors. The results showed that AAPR <0.58 was significantly related to higher pT stage and grade, concomitant variant histology, anemia and larger tumor size. Additionally, patients with a lower AAPR had an inferior survival outcomes than those with an AAPR ≥0.58 (all P < 0.001). Multivariate analysis suggested that the lower AAPR was also an independent risk factor for poor OS (HR 1.587, 95%CI: 1.185–2.126; P = 0.002), CSS (HR 1.746, 95%CI: 1.249–2.440; P = 0.001), and RFS (HR 1.337, 95%CI: 1.027–1.739; P = 0.031). Moreover, subgroup analysis demonstrated the lower AAPR was related to worse prognosis in high-grade UTUC patients; but in those with low-grade disease, no relationship between them was observed. In conclusion, our results found that the decreased AAPR was independently related to poor survival outcomes in UTUC patients. Using the AAPR for subclassification of high-grade UTUC seems to further identify a poor prognostic group and contribute to clinical decisions making.

The tumor microenvironment is associated with various tumor progressions, including cancer metastasis, immunosuppression, and tumor sustained growth. Tumor‐associated macrophages (TAMs) are considered an indispensable component of the tumor microenvironment, participating in the progression of tumor microenvironment remodeling and creating various compounds to regulate tumor activities. This study aims to observe enriched TAMs in tumor tissues during bladder cancer development, which markedly facilitated the proliferation of bladder cancer cells and promoted tumor growth in vivo. We determined that TAMs regulate tumor sustained growth by secreting type I collagen, which can activate the prosurvival integrin α2β1/PI3K/AKT signaling pathway. Furthermore, traditional chemotherapeutic drugs combined with integrin α2β1 inhibitor showed intensive anticancer effects, revealing an innovative approach in clinical bladder cancer treatment.