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Biology subjects

Parazzoli, D.

Publications and source records attributed to Parazzoli, D..

6 recordsLinked to original sources

Compressive stress couples calcium signalling to translational control and proteostasis

Compressive stress is a pervasive feature of solid tumour growth, and mechanical loading is known to activate calcium signalling through the mechanosensitive channel Piezo1. How this signal is converted into changes in cellular biosynthetic capacity, however, remains unclear. Here we show that compressive stress engages a Piezo1-dependent calcium checkpoint that reduces translational output with increasing mechanical load, independently of mTORC1 and the integrated stress response. Compression triggers Piezo1-dependent release of endoplasmic reticulum Ca2, activating eEF2 kinase to slow translational elongation. This same Ca2 signal drives mitochondrial remodelling and reactive oxygen species production, which reinforce translational repression through redox-dependent oxidation of the translation initiation factor eIF2{beta}. In parallel, Ca2-dependent AMPK activation triggers autophagic ribosome degradation, alleviating compression-induced macromolecular crowding and restoring cytoplasmic rheology despite sustained mechanical load. These mechanisms act at distinct levels of translational control: initiation, elongation and ribosome abundance; yet converge to limit biosynthetic output under compression. Consistent with a role for this pathway in tumour adaptation to mechanical constraint, eEF2K expression correlates with poor overall survival in a public PDAC patient dataset. Together, these findings define a mechano-metabolic checkpoint through which cells actively tune biosynthetic capacity to their physical environment.

cell biology↗

CDK12 controls transcription at damaged genes and prevents MYC-induced transcription-replication conflicts

Oncogene-induced replicative stress is a potent tumor-suppressive mechanism that must be kept in check for cancer cells to thrive. Thus, the identification of genes and pathways involved in replicative stress is key to understand cancer evolution and to identify prospective therapeutic targets. Here, we investigated factors that modulate replicative stress upon deregulation of the MYC oncogene. We identified the cyclin-dependent kinase CDK12 as selectively required to prevent transcription-replication conflicts and the activation of a cytotoxic DNA-damage response (DDR). At the mechanistic level, CDK12 was recruited to damaged genes by PARP-dependent DDR-signaling and elongation-competent RNAPII. Once recruited, CDK12 repressed transcription by preventing the association of CDK9 with RNAPII. Either loss or chemical inhibition of CDK12 led to DDR-resistant transcription at damaged genes. Genome-wide profiling revealed that loss of CDK12 exacerbated transcription-replication conflicts in MYC-overexpressing cells and led to the accumulation of double-strand DNA breaks (DSBs), occurring preferentially between early- replicating regions and transcribed genes, organized in a co-directional head-to-tail orientation. Overall, our data demonstrate that CDK12 protects genome integrity by repressing transcription of damaged genes, which is required for proper resolution of DSBs at oncogene-induced transcription-replication conflicts. This provides a rationale that explains both how CDK12 deficiency can promote tandem duplications of early-replicated regions during tumor evolution, and how CDK12 targeting can exacerbate replicative-stress in tumors.

cancer biology↗

Decellularized Normal and Tumor Scaffolds for Cancer Organoid Cultures as a Model of Colorectal Peritoneal Metastases

Peritoneal metastases (PM) from colorectal cancer (CRC) are associated with poor survival. The extracellular matrix (ECM) plays a fundamental role in modulating the homing of CRC metastases to the peritoneum. The mechanisms underlying the interactions between metastatic cells and the ECM, however, remain poorly understood and the number of in vitro models available for the study of the peritoneal metastatic process is limited. Here, we show that decellularized ECM of the peritoneal cavity allows the growth of organoids obtained from PM, favoring the development of three-dimensional nodules that maintain the characteristics of in vivo PM. Organoids preferentially grow on scaffolds obtained from neoplastic peritoneum, which are characterized by greater stiffness than normal scaffolds. A gene expression analysis of organoids grown on different substrates reflected faithfully the clinical and biological characteristics of the organoids. An impact of the ECM on the response to standard chemotherapy treatment for PM was also observed. SignificanceEvidence of the value of ex vivo 3D models obtained by combining patient-derived extracellular matrices depleted of cellular components and organoids to mimic the metastatic niche, to be used as a tool to develop new therapeutic strategies in a biologically relevant context, to personalize treatments and increase their efficacy.

cancer biology↗

The ubiquitin ligase Hecw controls oogenesis and neuronal homeostasis by promoting the liquid state of ribonucleoprotein particles

Specialised ribonucleoprotein (RNP) granules are a hallmark of germ cells. Among their main function is the spatial and temporal modulation of the activity of specific mRNA transcripts that allow specification of primary embryonic axes. While RNPs composition and role are well established, their regulation is poorly defined. Here, we demonstrate that Hecw, a newly identified Drosophila ubiquitin ligase, is a key modulator of RNPs in oogenesis. Loss of Hecw activity results in the formation of enlarged granules that transition from a liquid to a gel-like state. At the molecular level, Hecw depletion leads to reduced ubiquitination and activity of the translational repressor Fmrp, resulting in premature Orb expression/recruitment in nurse cells. In addition to defective oogenesis, flies lacking Hecw show neurodegenerative traits with premature aging and climbing defects due to neuronal loss that are linked to RNPs condensation. Our findings reveal an unprecedented function of ubiquitin in modulating RNP fluidity and activity.Competing Interest StatementThe authors have declared no competing interest.View Full Text

animal behavior and cognition↗

From injury to full repair: nerve regeneration and functional recovery in the common octopus, Octopus vulgaris

Spontaneous nerve regeneration in cephalopod molluscs occurs in a relative short time after injury, achieving functional recovery of the lost capacities. In particular, transection of the pallial nerve in the common octopus (Octopus vulgaris) determines loss and subsequent restoring of two functions fundamental for survival, i.e. breathing and skin patterning, the latter involved in communication between animals and concealing. The phenomena occurring after lesion have been investigated in a series of previous studies, but a complete analysis of the changes occurring at the level of the axons and the effects on animals appearance during the whole regenerative process is still missing. Our goal is to determine the course of events following injury. Our goal is to determine the course of events following injury, from impairment to full recovery.\n\nWe observed nerve regeneration, end-target re-innervation and functional reconnections between central brain and periphery, using the contralateral nerve in the same animal as internal control. The final architecture of the regenerated nervous tissue does not mirror the original structure, however functionality returns to match the phenotype of an intact octopus, and with no visible impact on the behaviour of the animal. This provides exceptional value to these findings for future studies.\n\nSummary statementHere we report events occurring after interruption of the peripheral neural circuitry in Octopus vulgaris, from the dramatic loss of normal functioning to full recovery.

neuroscience↗

Unjamming overcomes kinetic and proliferation arrest in terminally differentiated cells and promotes collective motility of carcinoma.

During wound repair, branching morphogenesis and carcinoma dissemination, cellular rearrangements are fostered by a solid-to-liquid transition known as unjamming. The biomolecular machinery behind unjamming, its physiological and clinical relevance remain, however, a mystery. Here, we combine biophysical and biochemical analysis to study unjamming in a variety of epithelial 2D and 3D collectives: monolayers, differentiated normal mammary cysts, spheroid models of breast ductal carcinoma in situ (DCIS), and ex vivo slices of orthotopically-implanted DCIS. In all cases, elevation of the small GTPase RAB5A sparks unjamming by promoting non-clathrin-dependent internalization of epidermal growth factor receptor that leads to hyper-activation of endosomally-confined ERK1/2 and phosphorylation of the actin nucleator WAVE2. Physically, activation of this pathway causes highly coordinated flocking of the cells, with striking rotational motion in 3D that eventually leads to matrix remodelling and collective invasiveness of otherwise jammed carcinoma. The identified endo-ERK1/2 pathway provides an effective switch for unjamming through flocking to promote epithelial tissues morphogenesis and carcinoma invasion and dissemination.

cancer biology↗