Association of mutation and expression of the brother of the regulator of imprinted sites (BORIS) gene with breast cancer progression

INTRODUCTION: The BORIS, 11 zinc-finger transcription factors, is a member of the cancer-testis antigen (CTA) family. It is mapped to chromosome number 20q13.2 and this region is genetically linked to the early onset of breast cancer. The current study analyzed the correlation between BORIS mutations and the expression of the protein in breast cancer cases. MATERIALS AND METHODS: A population-based study including a total of 155 breast cancer tissue samples and an equal number of normal adjacent tissues from Indian female breast cancer patients was carried out. Mutations of the BORIS gene were detected by polymerase chain reaction-single standard confirmation polymorphisms (PCR-SSCP) and automated DNA sequencing and by immunohistochemistry for BORIS protein expression were performed. The observed findings were correlated with several clinicopathological parameters to find out the clinical relevance of associations. RESULTS: Of all the cases 16.12% (25/155) showed mutations in the BORIS gene. The observed mutations present on codon 329 are missense, leading to Val\u3e Ile (G\u3eA) change on exon 5 of the BORIS gene. A significant association was observed between mutations of the BORIS gene and some clinicopathological features like nodal status (p = 0.013), estrogen receptor (ER) expression (p = 0.008), progesterone receptor (PR) expression (p = 0.039), clinical stage (p = 0.010) and menopausal status (p = 0.023). The protein expression analysis showed 20.64% (32/155) samples showing low or no expression (+), 34.19% (53/155) with moderate expression (++), and 45.17% (70/155) showing high expression (+++) of BORIS protein. A significant association was observed between the expression of BORIS protein and clinicopathological features like clinical stage (p = 0.013), nodal status (p = 0.049), ER expression (p = 0.039), and PR expression (p = 0.027). When mutation and protein expression were correlated in combination with clinicopathological parameters a significant association was observed in the category of high (+++) level of BORIS protein expression (p = 0.017). CONCLUSION: The BORIS mutations and high protein expression occur frequently in carcinoma of the breast suggesting their association with the onset and progression of breast carcinoma. Further, the BORIS has the potential to be used as a biomarker

Improving the treatment of intermediate risk (IR) B-cell precursor (BCP) acute lymphoblastic leukaemia

Most childhood ALL patients are stratified at diagnosis as Intermediate Risk (IR), although an unacceptably high proportion of these patients relapse. The lack of specific prognostic features makes it difficult to predict the response of IR patients to treatment. Recent progress in the development of patient-derived xenografts in immune-deficient mice represents an opportunity to improve relapse prediction in ALL patients. In this study, a pilot experiment was carried out to optimise the most appropriate engraftment strategy, which could stratify engraftment of samples from IR B cell Precursor (BCP) ALL patients according to clinical outcomes. High efficiency and quicker engraftment of IR ALL patient samples were observed in NSG mice and via the intravenous route of inoculation over NOD/SCID mice and the intrafemoral route. The response of xenografts to combination chemotherapy showed evidence of reflecting patient outcome. Validation of the capability of the optimised engraftment strategy to identify relapse in IR ALL patients using a larger patient cohort emphasised the role of drug treatment to discriminate between relapsed and non-relapsed IR ALL patients. Certainly, the time required for ALL patient samples to reach 25% engraftment in the peripheral blood of the drug-treated mice was the most significant parameter for predicting patient outcomes. Analysis of the ex vivo drug sensitivity of ALL xenografts revealed that the responses to vincristine and L-asparaginase reflected the in vivo responses to drug selection. Moreover, samples from one ALL xenograft selected in vivo with drug treatment showed increased resistance to vincristine ex vivo compared with the non drug-treated samples of the same xenograft. Microarray analysis of gene expression identified HSP90 as a potential target for reversing resistance, although the HSP90 inhibitor 17DMAG failed to enhance the inhibitory effect of vincristine ex vivo. Further analysis of gene expression revealed significant up-regulation of genes involved in dynamics and/or stability of the microtubule network in vincristine-resistant samples. In summary, engraftment of IR ALL samples under selective chemotherapy treatment could provide a clinical approach for upfront prediction of outcome in IR ALL patients, which could allow tailoring the intensity of treatment according to the risk of relapse within this patient subgroup

Liposome-Mediated Delivery of MERS Antigen Induces Potent Humoral and Cell-Mediated Immune Response in Mice

The advancements in the field of nanotechnology have provided a great platform for the development of effective antiviral vaccines. Liposome-mediated delivery of antigens has been shown to induce the antigen-specific stimulation of the humoral and cell-mediated immune responses. Here, we prepared dried, reconstituted vesicles (DRVs) from DPPC liposomes and used them as the vaccine carrier system for the Middle East respiratory syndrome coronavirus papain-like protease (DRVs-MERS-CoV PLpro). MERS-CoV PLpro emulsified in the Incomplete Freund’s Adjuvant (IFA-MERS-CoV PLpro) was used as a control. Immunization of mice with DRVs-MERS-CoV PLpro did not induce any notable toxicity, as revealed by the levels of the serum alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN) and lactate dehydrogenase (LDH) in the blood of immunized mice. Immunization with DRVs-MERS-CoV PLpro induced greater antigen-specific antibody titer and switching of IgG1 isotyping to IgG2a as compared to immunization with IFA-MERS-CoV PLpro. Moreover, splenocytes from mice immunized with DRVs-MERS-CoV PLpro exhibited greater proliferation in response to antigen stimulation. Moreover, splenocytes from DRVs-MERS-CoV PLpro-immunized mice secreted significantly higher IFN-γ as compared to splenocytes from IFA-MERS-CoV PLpro mice. In summary, DRVs-MERS-CoV PLpro may prove to be an effective prophylactic formulation to prevent MERS-CoV infection