Hyperthermia and associated changes in membrane fluidity potentiate P2X7 activation to promote tumor cell death

Extracellular ATP (eATP) accumulation within the tumor microenvironment (TME) has the potential to activate purinergic signaling. The eATP evoked signaling effects bolster antitumor immune responses while exerting direct cytotoxicity on tumor cells and vascular endothelial cells, mediated at least in part through P2X7 receptors. Approaches to augment purinergic signaling in TME e.g. by ectonucleotidase CD39 blockade, and/or boosting P2X7 functional responses, might be used as immunomodulatory therapies in cancer treatment. In this study, we delineated the translatable strategy of hyperthermia to demonstrate impacts on P2X7 responsiveness to eATP. Hyperthermia (40°C) was noted to enhance eATP-mediated cytotoxicity on MCA38 colon cancer cells. Increased membrane fluidity induced by hyperthermia boosted P2X7 functionality, potentiating pore opening and modulating downstream AKT/PRAS40/mTOR signaling events. When combined with cisplatin or mitomycin C, hyperthermia and eATP together markedly potentiate cancer cell death. Our data indicate that clinically tolerable hyperthermia with modulated P2X7-purinergic signaling will boost efficacy of conventional cancer treatments

NTPDase5/PCPH as a New Target in Highly Aggressive Tumors: A Systematic Review

The protooncogene PCPH was recently identified as being the ectonucleoside triphosphate diphosphohydrolase 5 (ENTPD5). This protooncogene is converted into an oncogene by a single base pair deletion, resulting in frame change and producing a premature stop codon, leading to a mutated protein (mt-PCPH) with only 27 kDa, which is much smaller than the original 47 kDa protein. Overexpression of the PCPH as well as the mutated PCPH increases the cellular resistance to stress and apoptosis. This is intriguing considering that the active form, that is, the oncogene, is the mutated PCPH. Several studies analyzed the expression of NTPDase5/mt-PCPH in a wide range of tumor cells and evaluated its role and mechanisms in cancer and other pathogenic processes. The main point of this review is to integrate the findings and proposed theories about the role played by NTPDase5/mt-PCPH in cancer progression, considering that these proteins have been suggested as potential early diagnostic tools and therapy targets

A Systematic Review of the Role of Purinergic Signalling Pathway in the Treatment of COVID-19

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global health concern. Three years since its origin, despite the approval of vaccines and specific treatments against this new coronavirus, there are still high rates of infection, hospitalization, and mortality in some countries. COVID-19 is characterised by a high inflammatory state and coagulation disturbances that may be linked to purinergic signalling molecules such as adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine (ADO), and purinergic receptors (P1 and P2). These nucleotides/nucleosides play important roles in cellular processes, such as immunomodulation, blood clot formation, and vasodilation, which are affected during SARS-CoV-2 infection. Therefore, drugs targeting this purinergic pathway, currently used for other pathologies, are being evaluated in preclinical and clinical trials for COVID-19. In this review, we focus on the potential of these drugs to control the release, degradation, and reuptake of these extracellular nucleotides and nucleosides to treat COVID-19. Drugs targeting the P1 receptors could have therapeutic efficacy due to their capacity to modulate the cytokine storm and the immune response. Those acting in P2X7, which is linked to NLRP3 inflammasome activation, are also valuable candidates as they can reduce the release of pro-inflammatory cytokines. However, according to the available preclinical and clinical data, the most promising medications to be used for COVID-19 treatment are those that modulate platelets behaviour and blood coagulation factors, mainly through the P2Y12 receptor

Activity and expression of ecto-nucleotide pyrophosphate/phosphodiesterases in a hepatic stellate cell line

7 p. : il.Nucleotides and nucleosides represent an important and ubiquitous class of molecules that interact with specific receptors, regulate a variety of activities within the liver, and play a role in the pathogenesis of hepatic fibrosis. Ecto-nucleotide pyrophosphatase/phosphodiesterases (E-NPPs) are ecto-enzymes that are located on the cell surface. NPP1, NPP2, and NPP3 (abbreviated as NPP1–3 hereafter) have been implicated in the hydrolysis of nucleotides; together with other ecto-nucleotidases, they control the events induced by extracellular nucleotides. We have identified and compared the expression of E-NPP family members in two different phenotypes of the mouse hepatic stellate cell line (GRX). In quiescent-like hepatic stellate cells (HSCs), E-NPP activity was significantly higher, NPP2 mRNA expression decreased and NPP3 mRNA increased. The differential NPP activity and expression in two phenotypes of GRX cells suggests that they are involved in the regulation of extracellular nucleotide metabolism in HSCs. However, the role of E-NPPs in the liver remains to be clarifie

Hierarchicky porézní biokeramika na bázi kompozitu geopolymer-hydroxyapatit jako nový biomateriál: Struktura, mechanické vlastnosti a hodnocení biokompatibility

V této studii byla technikou replik syntetizována porézní biokeramika s otevřenými buňkami na bázi geopolymerů. Složení se skládalo ze směsi metakaolinu (MK) a hydroxyapatitu (HA), které jsou v kombinaci vhodné pro použití v kostním tkáňovém inženýrství. Metakaolin je levný přírodní hlinitokřemičitan, který je známý pro své mechanické vlastnosti, zatímco hydroxyapatit vyniká svou biokompatibilitou, díky chemické struktuře podobné kostní matrix. Syntetizovaný geopolymer-hydroxyapatit (GMK-HA) byl po tepelném zpracování (HT) podroben tepelné úpravě (značen jako GMK-HA-HT). V práci byly vyhodnoceny chemicko-fyzikální vlastnosti keramických matric tj. chemické složení, porosita, morfologie, fázové složení, včetně buněčné interakce (ADSCs) s povrchem tohoto anorganického kompozitu.In this study, open-cell porous bioceramics based on geopolymers were synthesized by the replica technique. The composition consisted of a mixture of metakaolin (MK) and hydroxyapatite (HA) that combined are suitable for application in bone tissue engineering. Metakaolin is a cheap natural aluminosilicate known for its mechanical properties, while hydroxyapatite stands out for its biocompatibility due to the chemical structure similar to the bone matrix. After undergoing heat treatment (HT), the geopolymer-hydroxyapatite (GMK-HA) synthesized materials, referred to as GMK-HA-HT, presented pores in the range from 1 to 5 mm. In the compressive strength tests, they exhibited values between 1.18 and 2.9 MPa. These ranges of values signify a proper balance between mechanical strength and porosity. X-ray diffraction analysis showed phosphate and/or calcium crystalline phases in all heat-treated samples, indicating HA’s successful incorporation in the geopolymer structure. In vitro tests using human adipose-derived mesenchymal stem cells (ADSCs) were conducted to evaluate the biocompatibility of the synthesized materials. The extracts obtained from the GMK-HA-HT were found to be non-cytotoxic. ADSCs in contact with extracts have no morphological changes and when cultured on the GMK-HA-HT surface, the cells interacted with the scaffold and formed a monolayer. Here, for the first time, we provide insights into this new class of materials as a promising biomaterial candidate which could be associated with ADSCs to promote bone healing

New therapy of skin repair combining adipose-derived mesenchymal stem cells with sodium carboxymethylcellulose scaffold in a pre-clinical rat model.

Lesions with great loss of skin and extensive burns are usually treated with heterologous skin grafts, which may lead rejection. Cell therapy with mesenchymal stem cells is arising as a new proposal to accelerate the healing process. We tested a new therapy consisting of sodium carboxymethylcellulose (CMC) as a biomaterial, in combination with adipose-derived stem cells (ADSCs), to treat skin lesions in an in vivo rat model. This biomaterial did not affect membrane viability and induced a small and transient genotoxicity, only at the highest concentration tested (40 mg/mL). In a rat wound model, CMC at 10 mg/mL associated with ADSCs increased the rate of cell proliferation of the granulation tissue and epithelium thickness when compared to untreated lesions (Sham), but did not increase collagen fibers nor alter the overall speed of wound closure. Taken together, the results show that the CMC is capable to allow the growth of ADSCs and is safe for this biological application up to the concentration of 20 mg/mL. These findings suggest that CMC is a promising biomaterial to be used in cell therapy