Céline Delierneux

Summary: Despite the established role of mitochondria in tumorigenesis, the molecular mechanisms by which mitochondrial Ca ²⁺ (mtCa ²⁺ ) signaling regulates tumor growth and metastasis remain unknown. The crucial role of mtCa ²⁺ in tumorigenesis is highlighted by the altered expression of proteins mediating mtCa ²⁺ uptake and extrusion in cancer cells. Here, we demonstrate that expression of the mitochondrial Na ⁺ /Ca ²⁺ exchanger NCLX ( SLC8B1 ) is decreased in colorectal tumors and is associated with advanced-stage disease in patients. We reveal that downregulation of NCLX leads to mtCa ²⁺ overload, mitochondrial depolarization, mitophagy, and reduced tumor size. Concomitantly, NCLX downregulation drives metastatic spread, chemoresistance, the expression of epithelial-to-mesenchymal transition (EMT), hypoxia, and stem cell pathways. Mechanistically, mtCa ²⁺ overload leads to an increase in mitochondrial reactive oxygen species (mtROS) which activates HIF1α signaling supporting the metastatic behavior of tumor cells lacking NCLX. Our results reveal that loss of NCLX expression is a novel driver of metastatic progression, indicating that control of mtCa ²⁺ levels is a novel therapeutic approach in metastatic colorectal cancer. Highlights: The expression of NCLX is decreased in colorectal tumors and is associated with advanced-stage disease in patients. NCLX plays a dichotomous role in colorectal tumor growth and metastasis. NCLX downregulation causes mitophagy and reduced colorectal cancer tumor growth. NCLX downregulation induces stemness, chemoresistance and metastasis through mtCa ²⁺ /ROS/HIF1α signaling axis. Graphical Significance: Mitochondrial Ca ²⁺ (mtCa ²⁺ ) homeostasis is essential for cellular metabolism and growth and plays a critical role in cancer progression. mtCa ²⁺ uptake is mediated by an inner membrane protein complex containing the mitochondrial Ca ²⁺ uniporter (MCU). mtCa ²⁺ uptake by the MCU is followed by a ∼100-fold slower mtCa ²⁺ extrusion mediated by the inner mitochondrial membrane ion transporter, the mitochondrial Na ⁺ /Ca ²⁺ exchanger NCLX. Because NCLX is a slower transporter than the MCU, it is a crucial rate-limiting factor of mtCa ²⁺ homeostasis that cannot easily be compensated by another Ca ²⁺ transport mechanism. This represents the first study investigating the role of NCLX in tumorigenesis and metastasis. We demonstrate for the first time that colorectal cancers exhibit loss of NCLX expression and that this is associated with advanced-stage disease. Intriguingly, decreased NCLX function has a dichotomous role in colorectal cancer. Thus, we reveal that NCLX loss leads to reduced primary tumor growth and overall tumor burden in vivo . Yet, the consequential increases in mtCa ²⁺ elicit pro-survival, hypoxic and gene transcription pathways that enhance metastatic progression. This dichotomy is a well-established feature of chemoresistant and recurrent tumor cells including cancer stem cells. Moreover, the downstream changes elicited by NCLX loss are reminiscent of mesenchymal colorectal cancer subtypes that display poor patient survival. Our data indicate that the demonstrated changes to the mtCa ²⁺ /mtROS/HIF1α signaling axis elicited through the loss of NCLX are a key adaptation and driver of metastatic colorectal cancer.

Calcium (Ca²⁺) uptake into the mitochondria shapes cellular Ca²⁺ signals and acts as a key effector for ATP generation. In addition, mitochondria-derived reactive oxygen species (mROS), produced as a consequence of ATP synthesis at the electron transport chain (ETC), modulate cellular signaling pathways that contribute to many cellular processes. Cancer cells modulate mitochondrial Ca²⁺ ([Ca²⁺]m) homeostasis by altering the expression and function of mitochondrial Ca²⁺ channels and transporters required for the uptake and extrusion of mitochondrial Ca²⁺. Regulated elevations in [Ca²⁺]m are required for the activity of several mitochondrial enzymes, and this in turn regulates metabolic flux, mitochondrial ETC function and mROS generation. Alterations in both [Ca²⁺]m and mROS are hallmarks of many tumors, and elevated mROS is a known driver of pro-tumorigenic redox signaling, resulting in the activation of pathways implicated in cellular proliferation, metabolic alterations and stress-adaptations. In this review, we highlight recent studies that demonstrate the interplay between [Ca²⁺]m and mROS signaling in cancer.

Key Points: Soluble GPVI is elevated in patients with thermal injury with sepsis, and sGPVI levels augment severity score prediction of mortality. The GPVI ligand, fibrin, induces GPVI shedding without requirement for platelet activation or signaling

Vaccinia-H1 Related (VHR) dual-specificity phosphatase, or DUSP3, plays an important role in cell cycle regulation and its expression is altered in several human cancers. In mouse model, DUSP3 deletion prevents neo-angiogenesis and b-FGF-induced microvessel outgrowth. Considering the importance of angiogenesis in metastasis formation, our study aimed to investigate the role of DUSP3 in tumour cell dissemination. Using a Lewis Lung carcinoma (LLC) experimental metastasis model, we observed that DUSP3^-/- mice developed larger lung metastases than littermate controls. DUSP3^-/- bone marrow transfer to lethally irradiated DUSP3^+/+ mice was sufficient to transfer the phenotype to DUSP3^+/+ mice, indicating that hematopoietic cells compartment was involved in the increased tumour cell dissemination to lung tissues. Interestingly, we found a higher percentage of tumour-promoting Ly6C^int macrophages in DUSP3^-/- LLC-bearing lung homogenates that was at least partially due to a better recruitment of these cells. This was confirmed by 1) the presence of higher number of the Ly6B^hi macrophages in DUSP3^-/- lung homogenates and by 2) the better migration of DUSP3^-/- bone marrow sorted monocytes, peritoneal macrophages and bone marrow derived macrophages (BMDMs), compared to DUSP3^+/+ monocytes, macrophages and BMDMs, in response to LLC-conditioned medium. Our study demonstrates that DUSP3 phosphatase plays a key role in metastatic growth through a mechanism involving the recruitment of macrophages towards LLC-bearing lungs.

Platelets have been involved in both immune surveillance and host defense against severe infection. To date, whether platelet phenotype or other hemostasis components could be associated with predisposition to sepsis in critical illness remains unknown. The aim of this work was to identify platelet markers that could predict sepsis occurrence in critically ill injured patients. This single-center, prospective, observational, 7-month study was based on a cohort of 99 non-infected adult patients admitted to ICUs for elective cardiac surgery, trauma, acute brain injury, and post-operative prolonged ventilation and followed up during ICU stay. Clinical characteristics and severity score (SOFA) were recorded on admission. Platelet activation markers, including fibrinogen binding to platelets, platelet membrane P-selectin expression, plasma soluble CD40L, and platelet-leukocytes aggregates were assayed by flow cytometry at admission and 48 h later, and then at the time of sepsis diagnosis (Sepsis-3 criteria) and 7 days later for sepsis patients. Hospitalization data and outcomes were also recorded. Of the 99 patients, 19 developed sepsis after a median time of 5 days. These patients had a higher SOFA score at admission; levels of fibrinogen binding to platelets (platelet-Fg) and of D-dimers were also significantly increased compared to the other patients. Levels 48 h after ICU admission no longer differed between the two patient groups. Platelet-Fg % was an independent predictor of sepsis (P = 0.0031). By ROC curve analysis, cutoff point for Platelet-Fg (AUC = 0.75) was 50%. In patients with a SOFA cutoff of 8, the risk of sepsis reached 87% when Platelet-Fg levels were above 50%. Patients with sepsis had longer ICU and hospital stays and higher death rate. Platelet-bound fibrinogen levels assayed by flow cytometry within 24 h of ICU admission help identifying critically ill patients at risk of developing sepsis.