Biology subjects

Patriarca, E. J.

Publications and source records attributed to Patriarca, E. J..

3 recordsLinked to original sources

bioRxiv2026

Mechanobiology-guided drug repurposing identifies budesonide as an inhibitor of stiffness-induced PDAC aggressiveness

Pancreatic ductal adenocarcinoma (PDAC) develops within a desmoplastic and stiffened microenvironment that critically shapes tumor progression and therapeutic resistance, yet these features are not reproduced by conventional rigid plastic culture systems. Here, we leverage a tuneable bioengineered platform that mimics stromal stiffening to investigate how mechanical cues regulate PDAC cell behaviour and to identify pharmacological strategies that counteract stiffness-driven malignancy. We show that increasing matrix stiffness promotes key hallmarks of PDAC aggressiveness, including enhanced cell spreading, focal adhesions maturation, and cytoskeletal tension. Notably, we identify the glucocorticoid budesonide as a selective suppressor of stiffness-induced malignant phenotypes. Transcriptomic profiling reveals that budesonide counteracts stiffness-associated gene programs, prominently affecting pathways governing cytoskeletal dynamics, nuclear envelope organization, and YAP nucleocytoplasmic transport. Consistently, budesonide reduced force transmission to the nucleus, restoring nuclear wrinkling and constraining nuclear size and shape. These effects are mediated through both glucocorticoid receptor-dependent and -independent mechanisms, revealing a previously unrecognized mode of action. Together, our findings establish mechanical context as a critical determinant of PDAC vulnerability and identify budesonide as a candidate for therapeutic repurposing to target stiffness-driven cancer progression.

bioengineering↗

bioRxiv2024

Non-cell-autonomous control of gastruloid development by the lncRNA T-UCstem1 through DKK1-dependent modulation of WNT signalling

The role and the mechanisms of long non-coding RNA (lncRNAs) in early mammalian embryogenesis remain unclear; this is mostly due to the complexity of the regulatory mechanism of lncRNAs and the limited availability of early embryo samples. The emergence of stem cell-based models of early mammalian embryogenesis offers new opportunities to address these challenging questions. Here we use mouse gastruloids to investigate the role of an ultraconserved lncRNA, T-UCstem1, in the formation of the mammalian body plan. Combining morphological and immunofluorescence analysis with bulk and single cells transcriptomics, we provide unprecedented evidence that T-UCstem1 is a key regulator of gastruloid development and is required for the extension of the anteroposterior axis. Specifically, knock down of T-UCstem1 results in aberrant gastruloid development, which is characterized by altered spatiotemporal expression of the differentiation markers and persistence of pluripotency genes. Single-cell analysis reveals higher cellular heterogeneity in T-UCstem1 KD gastruloids. Notably, the presence of cell populations characterized by the co-expression of pluripotency and differentiation markers points to a key role of T-UCstem1 in establishment and maintenance of proper cellular identity. Mechanistically, we show that T-UCstem1 acts non-cell autonomously through Dickkopf-related protein 1 (DKK-1)-dependent modulation of WNT pathway. Our findings highlight a previously unexplored role for ultraconserved lncRNAs in gastruloid development and open the way for using gastruloids to dissect the functions of lncRNAs in early mammalian development.

developmental biology↗

bioRxiv2019

Gastruloid development competence discriminates different states of pluripotency between naïve and primed

Floating spheroidal aggregates (aggregomes) of mouse embryonic stem cells (mESCs) can develop into polarized/elongated organoids, namely gastruloids. Here we report a high-performing assay to measure gastruloids formation efficiency (GFE), i.e. the fraction of gastruloid-developing aggregomes. By exploiting this procedure, we provide morphological and molecular evidence that gastruloid development relies on Cripto. We also demonstrate that GFE decreases as pluripotency progresses from naive to primed state. Indeed, naive ESC-derived aggregomes efficiently elongate (GFE[≥]95%), while primed EpiSCs fail to aggregate and consequently to generate gastruloids (GFE=0%). Conversely, while early-primed EpiLCs properly aggregate, EpiLC-derived aggregomes are mostly abortive (GFE=0%). Unlike EpiLCs, L-Proline-treated ESCs (PiCs) generate productive aggregomes (GFE[≥]50%), which however begin to elongate earlier and generate smaller gastruloids that appear more differentiated. Like EpiLCs, PiCs are competent to differentiate into primordial germ cell-like cells (PGCLCs), suggesting that PiCs capture an EpiLC-like state with unique competence for both gastruloid formation and differentiation into PGCLCs. Thus we propose GFE assay as a simple and robust in vitro method to discriminate different phenotypic/functional states of the pluripotency continuum.

developmental biology↗