Inhibition of HIV-1 gene expression by retroviral vector-mediated small-guide RNAs that direct specific RNA cleavage by tRNase ZL

The tRNA 3′-processing endoribonuclease (tRNase Z or 3′ tRNase; EC 3.1.26.11) is an essential enzyme that removes the 3′ trailer from pre-tRNA. The long form (tRNase ZL) can cleave a target RNA in vitro at the site directed by an appropriate small-guide RNA (sgRNA). Here, we investigated whether this sgRNA/tRNase ZL strategy could be applied to gene therapy for AIDS. We tested the ability of four sgRNA-expression plasmids to inhibit HIV-1 gene expression in COS cells, using a transient-expression assay. The three sgRNAs guide inhibition of HIV-1 gene expression in cultured COS cells. Analysis of the HIV-1 mRNA levels suggested that sgRNA directed the tRNase ZL to mediate the degradation of target RNA. The observation that sgRNA was localized primarily in nuclei suggests that tRNase ZL cleaves the HIV-1 mRNA when complexed with sgRNA in this location. We also examined the ability of two retroviral vectors expressing sgRNA to suppress HIV-1 expression in HIV-1-infected Jurkat T cells. sgRNA-SL4 suppressed HIV-1 expression almost completely in infected cells for up to 18 days. These results suggest that the sgRNA/tRNase ZL approach is effective in downregulating HIV-1 gene expression

Increasing access to HIV testing for men who have sex with men in Japan using digital vending machine technology

Background: In Japan, most new HIV cases are reported amongst men who have sex with men (MSM); thus, there is an urgent need for further widespread testing of MSM. The use of Digital Vending Machines (DVM) in the UK offering HIV test kits targeting MSM show promising results. Digital Vending Machines could be useful to promote and increase the uptake of testing in Japan, although no studies have yet been conducted. We aimed to assess the acceptability and feasibility of distributing HIV test kits using DVMs exploring needs and concerns as well as preferred types of test kits and locations. Methods: Fifty-four individuals participated in workshops and meetings with a further 224 MSM answering a quantitative survey assessing HIV testing and prevention needs. Results: Amongst MSM who had never been tested, 73% showed willingness to purchase tests from DVMs. Responses were broadly positive about DVMs but there were concerns regarding being seen receiving test kits from the machines and linkage to confirmatory testing and appropriate care. Conclusions: Using DVMs to distribute HIV test kits in Japan was found to be both acceptable and feasible and may have the potential to increase access to testing for MSM. Future large-scale evaluation studies are required

Everywhere in Japan: an international approach to working with commercial gay businesses in HIV prevention

Summary In the UK and Japan, there is concern regarding rising rates of annual new HIV infections among Men who have Sex with Men (MSM). Whilst in the UK and Europe, gay businesses are increasingly recognized as being important settings through which to deliver HIV prevention and health promotion interventions to target vulnerable populations; in Japan such settings-based approaches are relatively underdeveloped. This article draws on qualitative data from a recently completed study conducted to explore whether it is feasible, acceptable and desirable to build on the recent European Everywhere project for adaptation and implementation in Japan. A series of expert workshops were conducted in Tokyo, Nagoya and Osaka with intersectoral representatives from Japanese and UK non-governmental organizations (NGOs), gay businesses, universities and gay communities (n = 46). Further discussion groups and meetings were held with NGO members and researchers from the Japanese Ministry of Health, Labour and Welfare's Research Group on HIV Prevention Policy, Programme Implementation and Evaluation among MSM (n = 34). The results showed that it is desirable, feasible and acceptable to adapt and implement a Japanese version of Everywhere. Such a practical, policy-relevant, settingsbased HIV prevention framework for gay businesses may help to facilitate the necessary scale up of prevention responses among MSM in Japan. Given the high degree of sexual mobility between countries in Asia, there is considerable potential for the Everywhere Project (or its Japanese variant) to be expanded and adapted to other countries within the Asia-Pacific region

Everywhere in Japan: An international approach to working with commercial gay businesses in HIV prevention

In the UK and Japan, there is concern regarding rising rates of annual new HIV infections among Men who have Sex with Men (MSM). Whilst in the UK and Europe, gay businesses are increasingly recognized as being important settings through which to deliver HIV prevention and health promotion interventions to target vulnerable populations; in Japan such settings-based approaches are relatively underdeveloped. This article draws on qualitative data from a recently completed study conducted to explore whether it is feasible, acceptable and desirable to build on the recent European Everywhere project for adaptation and implementation in Japan. A series of expert workshops were conducted in Tokyo, Nagoya and Osaka with intersectoral representatives from Japanese and UK non-governmental organizations (NGOs), gay businesses, universities and gay communities (n = 46). Further discussion groups and meetings were held with NGO members and researchers from the Japanese Ministry of Health, Labour and Welfare's Research Group on HIV Prevention Policy, Programme Implementation and Evaluation among MSM (n = 34). The results showed that it is desirable, feasible and acceptable to adapt and implement a Japanese version of Everywhere. Such a practical, policy-relevant, settings-based HIV prevention framework for gay businesses may help to facilitate the necessary scale up of prevention responses among MSM in Japan. Given the high degree of sexual mobility between countries in Asia, there is considerable potential for the Everywhere Project (or its Japanese variant) to be expanded and adapted to other countries within the Asia-Pacific region

Real-world status of treatment for lymphoid neoplasms developed during the course of myeloproliferative neoplasms in Japan

ABSTRACTObjectives: Patients with myeloproliferative neoplasms (MPNs) are at higher risk of developing secondary malignancies. In this study, we focused on patients with MPNs that complicated lymphoid neoplasms. To analyze the real-world status of lymphoid neoplasm treatment in patients with pre-existing MPNs in Japan, we conducted a multicenter retrospective study.Methods: Questionnaires were sent to collect the data on patients who were first diagnosed with either polycythemia vera, essential thrombocythemia or myelofibrosis and who later were complicated with lymphoid neoplasms defined as malignant lymphoma, multiple myeloma, or chronic lymphocytic leukemia/small cell lymphoma.Results: Twenty-four patients with MPNs complicated by lymphoid neoplasms were enrolled (polycythemia vera, n = 8; essential thrombocythemia, n = 14; and primary myelofibrosis, n = 2). Among these, diffuse large B-cell lymphoma (DLBCL) was the most frequently observed (n = 13, 54.1%). Twelve (92.3%) of the patients with DLBCL received conventional chemotherapy. Among these 12 patients, regarding cytoreductive therapy for MPNs, 8 patients stopped treatment, one continued treatment, and two received a reduced dose. Consequently, most patients were able to receive conventional chemotherapy for DLBCL with a slightly higher dose of granulocyte colony-stimulating factor support than usual without worse outcomes. All 3 patients with multiple myeloma received a standard dose of chemotherapy.Conclusion: Our data indicate that if aggressive lymphoid neoplasms develop during the course of treatment in patients with MPNs, it is acceptable to prioritize chemotherapy for lymphoma