Biology subjects

Romano, E.

Publications and source records attributed to Romano, E..

4 recordsLinked to original sources

bioRxiv2026

In situ programming of intratumoural stem cell-like memory CD8+ T cells enables durable antitumour immunity in immunosuppressive tumours

Tumours with an immunosuppressive microenvironment often respond poorly to radiotherapy (RT) and immune checkpoint inhibitors, and durable clinical responses remain rare. These failures are not solely explained by insufficient immune activation, but instead reflect an inability of current therapies to establish CD8 T-cell infiltration and states required for long-term tumour control. Although stem cell-like memory CD8 T cells (Tscm) are critical for durable antitumour immunity, they are typically rare within solid tumours, and strategies to selectively augment the expansion of intratumoural Tscm within immunosuppressive tumours are lacking. Here, we show that intratumoural immune modulation via pattern recognition receptor (PRR) activation, using a double-stranded RNA-based agonist (BO-112), promotes Tscm expansion and, in combination with radiation, enhances antitumour immunity in poorly immunogenic lung and bladder cancer models. PRR activation enabled selective in situ expansion of intratumoural Tscm, rather than global CD8 T-cell activation. Therapeutic efficacy required pre-existing intratumoural CD8 T cells and was associated with qualitative reprogramming of the immune landscape, including depletion of exhausted CD8 T cells and regulatory T cells. Transcriptomic analyses showed that PRR activation preferentially enriched programmes associated with Tscm formation and self-renewal, whereas durable effector differentiation emerged only following combined RT-PRR agonist treatment, consistent with antigen-driven execution of the expanded progenitor pool. A Tscm-associated gene signature derived from these preclinical datasets correlated with improved survival across independent human lung and bladder cancer cohorts. Together, our data identify intratumoural Tscm as a therapeutically targetable immune-control axis and establish in situ programming of Tscm as a route to durable antitumour immunity.

cancer biology↗

bioRxiv2025

Analysis of intratumoral immune heterogeneity reveals spatial organisation of immunosuppression in breast cancer

Recent advances in single-cell transcriptomics have illuminated intratumoral heterogeneity (ITH) in breast cancer, yet the spatial organization and functional states of immune populations within luminal tumors remain poorly understood. Here, we generated a spatially and clonally resolved single-cell immune atlas of early-stage luminal breast cancer by integrating paired single-cell RNA/TCR sequencing with multiplex immunohistochemistry across three spatially distinct tumor regions: central (core-proximal), intermediate and peripheral (invasive margin-proximal), in five patients. Peripheral regions are enriched in naive CD4 T cells, monocytes, dendritic cells, and memory CD8 T cells with bystander properties, alongside TCF1+ exhausted progenitors (Tpex), and displayed elevated antigen presentation and tertiary lymphoid structures (TLS) signatures. In contrast, central regions harbor macrophages, clonally expanded regulatory T cells (Tregs), and two subsets of terminally exhausted CD8 T cells (Tex), one tissue-resident and one circulating, both expressing tumor-reactive features. Clonal overlap was observed between peripheral Tpex and central, tissue-resident Tex cells, revealing a spatially confined differentiation trajectory toward exhaustion within the tumor microenvironment. Central T cells also upregulate TGF-{beta} signaling and metabolic pathways including fatty acid oxidation and oxidative phosphorylation, consistent with adaptation to an immunosuppressive microenvironment. Together, these findings reveal a spatially structured immune landscape in luminal breast cancer, marked by progressive immunosuppression from the tumor margin to the core, providing a framework for future region-specific immunotherapeutic strategies.

immunology↗

bioRxiv2024

Mitochondrial sites of contact with the nucleus aid in chemotherapy evasion of glioblastoma cells

Glioblastoma (GBM) is the most common form of a malignant primary brain tumour in adults for which therapeutic options are minimal. The rapid onset of the resistance mechanisms against the chemotherapeutic agent Temozolomide (TMZ), the first line of pharmacological care for patients, prevents the long-term validity of this approach. The underpinning biology for this remains poorly understood thus compromising the efficacy of this approach. The Translocator Protein (TSPO) is an 18kDa ubiquitous cholesterol-binding molecule on the outer membrane of mitochondria (OMM). Upregulated in cancers TSPO is required to form contacts between mitochondria and the nucleus termed: Nucleus Associated Mitochondria (NAM). In GBM tissues as well as in 2D and 3D cell cultures we assayed patterns of TSPO expression (i), autophagy/mitophagy (ii), transcription factors (iii) and susceptibility to TMZ-induced demise (iv). Confocal and ultrastructural imaging detailed the organization and redistribution of the mitochondrial network (v). Our findings show that TMZ exploits mitochondria via TSPO to aid the formation of NAM which couples the expression of the nuclear transcription factor Sterol regulatory element-binding transcription factor 1 (SREBP1) and the stabilization of YAP/TAZ. Pharmacological modulation of TSPO counteracts all the above and re-instates susceptibility to TMZ-induced demise. NAM is therefore proposed as a variable in the engagement and execution of pro-survival mechanisms in GBM thus offering a means to both insight into the pathophysiology of this disease and offer novel therapeutic strategies. Key PointsO_LITMZ exploits TSPO to curb mitochondrial quality control in glioblastoma cells. C_LIO_LITMZ-mediated MRR is associated with the relocation of mitochondria to the nucleus and modulation of transcriptional factors involved in cholesterol metabolism and adaptation to aggressive growth. C_LIO_LITSPO represents a pharmacological target to revert chemoresistance in glioblastoma cells. C_LI Importance of the StudyThis study elucidates a mitochondrion-driven mechanism of chemoresistance in human glioblastoma cells, which depends on the mitochondrial translocator protein TSPO. The administration of TSPO ligands restores susceptibility to TMZ by influencing the dynamics of transcriptional factors associated with cholesterol metabolism and mechanical transduction.

cancer biology↗

bioRxiv2020

An inducible AraC that responds to blue light instead of arabinose

In Escherichia coli, the operon responsible for the catabolism of L-arabinose is regulated by the dimeric DNA-binding protein AraC. In the absence of L-arabinose, AraC binds to the distal I1 and O2 half-sites, leading to repression of the downstream PBAD promoter. In the presence of the sugar, the dimer changes conformation and binds to the adjacent I1 and I2 half-sites, resulting in the activation of PBAD. Here we engineer blue light-inducible AraC dimers in Escherichia coli (BLADE) by swapping the dimerization domain of AraC with blue light-inducible dimerization domains. Using BLADE to overexpress proteins important for cell shape and division site selection, we reversibly control cell morphology with light. We demonstrate the exquisite light responsiveness of BLADE by employing it to create bacteriographs with an unprecedented quality. We then employ it to perform a medium-throughput characterization of 39 E. coli genes with poorly defined or completely unknown function. Finally, we expand the initial library and create a whole family of BLADE transcription factors (TFs), which we characterize using a novel 96-well light induction setup. Since the PBAD promoter is commonly used by microbiologists, we envisage that the BLADE TFs will bring the many advantages of optogenetic gene expression to the field of microbiology.

microbiology↗