Arejas Uzgiris

A Novel Strategy for Detection and Enumeration of Circulating Rare Cell Populations in Metastatic Cancer Patients Using Automated Microfluidic Filtration and Multiplex Immunoassay

Size selection via filtration offers an antigen-independent approach for the enrichment of rare cell populations in blood of cancer patients. We evaluated the performance of a novel approach for multiplex rare cell detection in blood samples from metastatic breast (n = 19) and lung cancer patients (n = 21), and healthy controls (n = 30) using an automated microfluidic filtration and multiplex immunoassay strategy. Captured cells were enumerated after sequential staining for specific markers to identify circulating tumor cells (CTCs), circulating mesenchymal cells (CMCs), putative circulating stem cells (CSCs), and circulating endothelial cells (CECs). Preclinical validation experiments using cancer cells spiked into healthy blood demonstrated high recovery rate (mean = 85%) and reproducibility of the assay. In clinical studies, CTCs and CMCs were detected in 35% and 58% of cancer patients, respectively, and were largely absent from healthy controls (3%, p = 0.001). Mean levels of CTCs were significantly higher in breast than in lung cancer patients (p = 0.03). Fifty-three percent (53%) of cancer patients harbored putative CSCs, while none were detectable in healthy controls (p<0.0001). In contrast, CECs were observed in both cancer and control groups. Direct comparison of CellSearch® vs. our microfluidic filter method revealed moderate correlation (R2 = 0.46, kappa = 0.47). Serial blood analysis in breast cancer patients demonstrated the feasibility of monitoring circulating rare cell populations over time. Simultaneous assessment of CTCs, CMCs, CSCs and CECs may provide new tools to study mechanisms of disease progression and treatment response/resistance.

Infection of human uterine fibroblasts by Zika virus in vitro: implications for viral transmission in women

Zika virus (ZIKV) is a single-stranded RNA arbovirus belonging to the Flavivirus genus. Similar to other zoonotic arboviruses, ZIKV is transmitted by the Aedes mosquito. It causes Zika fever, a usually non-fatal condition with symptoms including headache, fever, malaise, maculopapular rash, joint pains, and conjunctivitis. Recently, it was revealed that ZIKV may have additional pathogenic effects on pregnant women and their fetuses. Reports from South America suggest that ZIKV infection during pregnancy causes fetal microcephaly, which has negative impacts up to and including death. In addition to mosquito-borne transmission, it has been reported that ZIKV may be transmitted through heterosexual intercourse, with concomitant implications for global women’s health. This is the first example of a mosquito-borne arbovirus that may also utilize a sexual route of transmission. This knowledge has initiated a massive effort by the translational science community to develop more effective detection methods and treatment strategies. Currently there are no approved vaccines or drugs for preventing or treating ZIKV infection or ZIKV-induced pathologies. Recent diagnostics development efforts have focused on ELISA and highly sensitive quantitative reverse transcription PCR