The Stress Response of the Holothurian Central Nervous System: A Transcriptomic Analysis.

Injury to the central nervous system (CNS) results in permanent damage and lack of function in most vertebrate animals, due to their limited regenerative capacities. In contrast, echinoderms can fully regenerate their radial nerve cord (RNC) following transection, with little to no scarring. Investigators have associated the regenerative capacity of some organisms to the stress response and inflammation produced by the injury. Here, we explore the gene activation profile of the stressed holothurian CNS. To do this, we performed RNA sequencing on isolated RNC explants submitted to the stress of transection and enzyme dissection and compared them with explants kept in culture for 3 days following dissection. We describe stress-associated genes, including members of heat-shock families, ubiquitin-related pathways, transposons, and apoptosis that were differentially expressed. Surprisingly, the stress response does not induce apoptosis in this system. Other genes associated with stress in other animal models, such as hero proteins and those associated with the integrated stress response, were not found to be differentially expressed either. Our results provide a new viewpoint on the stress response in the nervous system of an organism with amazing regenerative capacities. This is the first step in deciphering the molecular processes that allow echinoderms to undergo fully functional CNS regeneration, and also provides a comparative view of the stress response in other organisms.

Metabolic diversity within breast cancer brain-tropic cells determines metastatic fitness.

HER2+ breast cancer patients are presented with either synchronous (S-BM), latent (Lat), or metachronous (M-BM) brain metastases. However, the basis for disparate metastatic fitness among disseminated tumor cells of similar oncotype within a distal organ remains unknown. Here, employing brain metastatic models, we show that metabolic diversity and plasticity within brain-tropic cells determine metastatic fitness. Lactate secreted by aggressive metastatic cells or lactate supplementation to mice bearing Lat cells limits innate immunosurveillance and triggers overt metastasis. Attenuating lactate metabolism in S-BM impedes metastasis, while M-BM adapt and survive as residual disease. In contrast to S-BM, Lat and M-BM survive in equilibrium with innate immunosurveillance, oxidize glutamine, and maintain cellular redox homeostasis through the anionic amino acid transporter xCT. Moreover, xCT expression is significantly higher in matched M-BM brain metastatic samples compared to primary tumors from HER2+ breast cancer patients. Inhibiting xCT function attenuates residual disease and recurrence in these preclinical models.

Dedifferentiation of radial glia-like cells is observed in in vitro explants of holothurian radial nerve cord.

BACKGROUND: Among animal phyla, some of the least studied nervous systems are those of the phylum Echinodermata. Part of the problem lies in that most of their nervous components are embedded in the body wall that has calcareous skeletal components. NEW METHOD: We have developed a novel technique for the successful isolation of the radial nerve cords (RNCs) and an in vitro system where the isolated RNCs can be cultured and are amenable to experimental manipulation. Here we use this system to isolate the RNC of the sea cucumber Holothuria glaberrima as a way to extend our studies on its regeneration capabilities. RESULTS: The RNCs can be isolated from the surrounding tissues by collagenase treatment. The explants obtained following enzymatic dissociation can be kept in culture for up to 2 weeks. Histological and immunohistochemical studies show that the explants maintain a stable number of cells with little proliferation or apoptosis throughout the culture incubation period. The main change observed in RNCs in vitro is a progressive dedifferentiation of radial glia-like cells. This dedifferentiation corresponds to the first step in the regeneration response to injury that has been described in vivo. COMPARISON WITH EXISTING METHODS: There are no existing methods to isolate and culture echinoderm radial nerve cord. CONCLUSIONS: The described protocol provides a unique tool to obtain easily accessible RNC from holothurians to perform cellular, biochemical, and genomic experiments in the echinoderm nervous system without interference of adjacent tissues. The technique provides a unique opportunity to study the dedifferentiation response associated with the regeneration of the nervous system in echinoderms.