Karki, A. | bioRxiv Science

bioRxiv2021

DNAJB1-PRKACA in HEK293T cells induces LINC00473 overexpression that depends on PKA signaling

Fibrolamellar carcinoma (FLC) is a primary liver cancer that most commonly arises in adolescents and young adults in a background of normal liver tissue and has an poor prognosis due to lack of effective chemotherapeutic agents. The DNAJB1-PRKACA gene fusion (DP) has been reported in the majority of FLC tumors, however its oncogenic mechanisms remain unclear. Given the paucity of cellular models, in particular FLC tumor cell lines, we hypothesized that engineering the DP fusion gene in HEK293T cells would provide insight into the cellular effects of the fusion gene. We used CRISPR/Cas9 to engineer HEK293T clones expressing DP fusion gene (HEK-DP) and performed transcriptomic, proteomic, and mitochondrial studies to characterize this cellular model. Proteomic analysis of DP interacting partners identified mitochondrial proteins as well as proteins in other subcellular compartments. HEK-DP cells demonstrated significantly elevated mitochondrial fission, which suggests a role for DP in altering mitochondrial dynamics. Transcriptomic analysis of HEK-DP cells revealed a significant increase in LINC00473 expression, similar to what has been observed in primary FLC samples. LINC00473 overexpression was reversible with siRNA targeting of PRKACA as well as pharmacologic targeting of PKA and Hsp40 in HEK-DP cells. Therefore, our model suggests that LINC00473 is a candidate marker for DP activity.

cancer biology↗

bioRxiv2020

In-stream habitat availability for river dolphins in response to flow: use of ecological integrity to manage river flows

Population decline and extinction risk of riverine cetaceans are primarily associated with flow alteration. Previous studies predominantly highlighted maintenance of adequate flow for low water seasons when habitats contract and the risk of local extinction escalates. Although riverine cetaceans are sensitive to reduction in river flow, no studies quantify the relationships between flow and ecology of riverine cetaceans to mitigate the potential adverse impacts of flow alteration. We quantify the relationships between flow and the ecology of river cetaceans with reference to Ganges River dolphins (GRD; Platanista gangetica gangetica) usable area availability (AWS) for the low water season at wider flows (50-575 m3/s) at finer spatial and temporal scales. This study reveals that distribution of usable area to GRD highly regulated by the adequate flow level as the interaction of appropriate velocity and depth at particular geomorphic that likely offer energetically efficient modes of locomotion to increase surfacing behavior efficiency, suggesting the hydro-physical environment as a major determinant of river cetaceans distribution and abundance. Flow and AWS relationships indicate that the flow during dry season negatively contributed to AWS, whereas that of pre-monsoon maximized the AWS, suggesting that modifying flow regimes does alter in-stream habitats at varying spatial scales and may influence life-history strategies. Substantial fragmentation in suitable pool availability and loss of longitudinal connectivity exhibited by dry season flow suggested a higher risk of adverse biological effects during the dry season, which may reduce population viability by reducing survivorship and reproduction failure. Owing to riverine cetaceans dependence on the attribute of freshwater flow, they can be expected to be more affected by flow regulations as interactive effects. Considering the seasonal effects and changes in the availability of usable areas by flow alteration, adopting effective habitat retention plans by water-based development projects appears critical to avoid further ecological risks in aquatic species conservation. Identifying priority riverscapes for river cetaceans and prioritizing investment opportunities is an essential first step towards effective riverine cetacean conservation.

ecology↗