No abstract

Despite current treatment regimens, heart failure remains the leading cause of morbidity and mortality in the developed world due to the limited capacity of adult mammalian ventricular cardiomyocytes to divide and replace ventricular myocardium lost from ischemia-induced infarct1,2. As a result, there is great interest to identify potential cellular sources and strategies to generate new ventricular myocardium3. Past studies have shown that lower vertebrate and early postnatal mammalian ventricular cardiomyocytes can proliferate to help regenerate injured ventricles4–6; however, recent studies have suggested that additional endogenous cellular sources may contribute to this overall ventricular regeneration3. Here, we have developed in the zebrafish a combination of fluorescent reporter transgenes, genetic fate-mapping strategies, and a ventricle-specific genetic ablation system to discover that differentiated atrial cardiomyocytes can transdifferentiate into ventricular cardiomyocytes to contribute to zebrafish cardiac ventricular regeneration. Using in vivo time-lapse and confocal imaging, we monitored the dynamic cellular events during atrial-to-ventricular cardiomyocyte transdifferentiation to define intermediate cardiac reprogramming stages. Importantly, we observed that Notch signaling becomes activated in the atrial endocardium following ventricular ablation, and discovered that inhibiting Notch signaling blocked the atrial-to-ventricular transdifferentiation and cardiac regeneration. Overall, these studies not only provide evidence for the plasticity of cardiac lineages during myocardial injury, but more importantly reveal an abundant new potential cardiac resident cellular source for cardiac ventricular regeneration.

Summary1. Photographic mark-recapture is a cost-effective, non-invasive way to study populations. However, to efficiently apply photographic mark-recapture to large populations, computer software is needed for image manipulation and pattern matching. 2.We created an open-source application for the storage, pattern extraction and pattern matching of digital images for the purposes of mark-recapture analysis. The resulting software package is a stand-alone, multiplatform application implemented in Java. Our program employs the Scale Invariant Feature Transform (SIFT) operator that extracts distinctive features invariant to image scale and rotation. 3. We applied this system to a population of Masai giraffe (Giraffa camelopardalis tippelskirchi) in the Tarangire Ecosystem in northern Tanzania. Over 1200 images were acquired in the field during three primary sampling periods between September 2008 and December 2009. The pattern information in these images was extracted and matched resulting in capture histories for over 600 unique individuals. 4. Estimated error rates of the matching system were low based on a subset of test images that were independently matched by eye. 5. Encounter histories were subsequently analysed with open population models to estimate apparent survival rates and population size. 6. This new open-access tool allowed photographic mark-recapture to be applied successfully to this relatively large population.

Sorafenib is the standard treatment for advanced hepatocellular carcinoma (HCC). However, the development of drug resistance is common. By using genome-wide CRISPR/Cas9 library screening, we identify phosphoglycerate dehydrogenase (PHGDH), the first committed enzyme in the serine synthesis pathway (SSP), as a critical driver for Sorafenib resistance. Sorafenib treatment activates SSP by inducing PHGDH expression. With RNAi knockdown and CRISPR/Cas9 knockout models, we show that inactivation of PHGDH paralyzes the SSP and reduce the production of αKG, serine, and NADPH. Concomitantly, inactivation of PHGDH elevates ROS level and induces HCC apoptosis upon Sorafenib treatment. More strikingly, treatment of PHGDH inhibitor NCT-503 works synergistically with Sorafenib to abolish HCC growth in vivo. Similar findings are also obtained in other FDA-approved tyrosine kinase inhibitors (TKIs), including Regorafenib or Lenvatinib. In summary, our results demonstrate that targeting PHGDH is an effective approach to overcome TKI drug resistance in HCC.