Although the combination of gene therapy and virotherapy for cancer therapy has obtained some encouraging results in vitro and in vivo over the past few years, some improvements of the vectors are still urgently needed to enhance their therapeutic effects for cancers. In order to maximize the anti-cancer activities of conditionally replicating adenoviral vectors (CRAd) vector, we for the first time generated a novel CRAd vector by inserting an expression cassette between E4 and the fiber using homologous recombination and tested this vector in melanoma cancer therapy. By using this novel vector we expressed two therapeutic transgenes, IL-24 and arresten, inserted in E1 and the region between E4 and the fiber, respectively, to test the melanoma inhibitory activities of this oncolytic virus in vitro and in vivo. The two therapeutic transgenes were successfully expressed by the novel CRAd, which was confirmed by western blotting. We then showed that this novel CRAd vector significantly suppressed the tumor growth of melanoma in vitro and in vivo compared with the control by inducing apoptosis and inhibiting angiogenesis. The novel CRAd vector generated in this study holds promise for developing more effective therapeutics for not only melanoma but also other common cancers.

The transduction efficiency of Ad (adenovirus) depends, to some extent, on the expression level of CAR (coxsackievirus and Ad receptor) of a target cell. The low level of CAR on the cell surface is a potential barrier to efficient gene transfer. To overcome this problem, PTD.AdeGFP (where eGFP is enhanced green fluorescent protein) was constructed by modifying the HI loop of Ad5 (Ad type 5) fibre with the Tat (trans-activating) PTD (protein transduction domain) derived from HIV. The present study showed that PTD.AdeGFP significantly improved gene transfer to multiple cell types deficient in expression of CAR. The improvement in gene transfer was not the result of charge-directed binding between the virus and the cell surface. Although PTD.AdeGFP formed aggregates, it infected target cells in a manner different from AdeGFP aggregates precipitated by calcium phosphate. In addition, PTD.AdeGFP was able to transduce target cells in a dynamin-independent pathway. The results provide some new clues as to how PTD.AdeGFP infects target cells. This new vector would be valuable in gene-function analysis and for gene therapy in cancer.

Progranulin (PGRN) encodes a 68.5-kDa secreted growth factor that is composed of seven and a half tandem repeats of a 12-cysteine granulin motif. PGRN is expressed in many tissues and has a role in mediating development, wound repair, inflammation, and tumorigenesis. Mutations leading to a loss of function in PGRN are the most common cause of familial frontotemporal lobar degeneration with TDP-43-positive inclusions (FTLD-TDP). In this study, we established hybridoma cell lines producing antibodies against human PGRN. Murine monoclonal antibodies (MAbs) against human PGRN were generated by using purified eukaryotic recombinant PGRN-6His fusion protein as immunogen. A panel of seven monoclonal antibodies was obtained after the preliminary screening by indirect enzyme-linked immunosorbent assay (ELISA), the data of which was confirmed by Western blot analysis and immunocytochemistry. By using constructs expressing a series of C- and N-terminal truncations, and single domains of PGRN, the particular domains recognized by MAbs were also identified. Domain-specific anti-PGRN MAbs will be an essential tool for investigating the role of PGRN in normal physiological or pathological conditions.

FAM76B has been reported to be a nuclear speckle localized protein with unknown function. In this study, FAM76B was first demonstrated to inhibit the NF-κB-mediated inflammatory pathway by affecting the translocation of hnRNPA2B1in vitro. We further showed that FAM76B suppressed inflammation by regulating the NF-κB pathwayin vivousing a traumatic brain injury (TBI) model in FAM76B knockout mice. Lastly, FAM76B was shown to interact with hnRNPA2B1 in human tissues taken from patients with acute, organizing, and chronic TBI, and with different neurodegenerative diseases. The results suggested that FAM76B mediates neuroinflammation by influencing the translocation of hnRNPA2B1in vivoduring TBI repair and neurodegenerative diseases. In summary, we for the first time demonstrated the role of FAM76B in regulating inflammation and further showed that FAM76B could regulate the NF-κB-mediated inflammatory pathway by affecting hnRNPA2B1 translocation, which provides new information for studying the mechanism of inflammation regulation.