Biology subjects

de la Roche, M.

Publications and source records attributed to de la Roche, M..

7 recordsLinked to original sources

bioRxiv2024

Angiogenic and Immune Predictors of Neoadjuvant Axitinib Response in Renal Cell Carcinoma with Venous Tumour Thrombus

Venous tumour thrombus (VTT), where the primary tumour invades the renal vein and inferior vena cava, affects 10-15% of renal cell carcinoma (RCC) patients. Curative surgery for VTT is high-risk, but neoadjuvant therapy may improve outcomes. The NAXIVA trial demonstrated a 35% VTT response rate after 8 weeks of neoadjuvant axitinib, a VEGFR-directed therapy. However, understanding non-response is critical for better treatment. We conducted a multiparametric investigation of samples collected during NAXIVA using digital pathology, flow cytometry, plasma cytokine profiling and RNA sequencing. Responders had higher baseline microvessel density and increased induction of VEGF-A and PlGF during treatment. A multi-modal machine learning model integrating features predicted response with an AUC of 0.868, improving to 0.945 when using features from week 3. Key predictive features included plasma CCL17 and IL-12. These findings may guide future treatment strategies for VTT, improving the clinical management of this challenging scenario. One Sentence SummaryA comprehensive multiparametric assessment of the effect of neoadjuvant axitinib in renal cell carcinoma patients with venous tumour thrombus was performed on tissue and peripheral blood, including an integrative machine learning model, which identified both angiogenic and immune determinants of response to therapy.

cancer biology↗

bioRxiv2024

Hedgehog signaling controls cytotoxic T cell migration in the tumour microenvironment.

Cytotoxic T lymphocytes effectively eliminate cancer cells. Their abundance in the tumour microenvironment is one of the strongest pan-cancer predictors of clinical response. Here, we show that Hedgehog (Hh) signaling regulates T cell migration into tumours. Using conditional knockout mouse models of central Hh signaling components Ihh, Smo and Gli1 in CD8 T cells, we show that Smo deletion greatly impairs the anti-tumour response in vivo due to diminished CD8 T cell migration into the tumour microenvironment. The migration defect is mediated exclusively by Smo, both in in vivo cancer models and in vitro migration assays. This effect is independent of the canonical Hh pathway and relies on the GPCR function of Smo to regulate the migration of murine and human CD8 T cells via RhoA. Hh signaling is critical during embryonic development and adult stem cell homeostasis, but is also amplified in multiple cancer types. Hh inhibitors targeting SMO have been clinically-approved and shown efficacy in the treatment of Hh-driven basal cell carcinoma and medulloblastoma but have failed in clinical trials in other solid cancers with upregulated Hh signaling. We demonstrate that SMO inhibitors specifically decrease CD8 T cell migration into the tumour microenvironment, both in murine cancer models and resected BCCs from patients treated with the SMO inhibitor vismodegib, providing the first mechanistic explanation as to why Hh inhibitors have failed in solid cancers. Our data establishes a novel link between Hh inhibition in vivo and the anti-tumour immune response and reveals a fundamental mechanism controlling T cell migration. The work provides the basis for improved Hh targeting approaches in the clinic and new entry points into enhancing migration in T cell therapies.

immunology↗

bioRxiv2022

LGR5 targeting molecules as therapeutic agents for multiple cancer types

Leucine-rich repeat-containing G-protein receptor 5 (LGR5) has been characterised as a stem cell and cancer stem cell marker. Previous analyses of LGR5 transcript levels indicate high level expression discriminates malignancies such as colorectal cancer (CRC) and pre-B acute lymphoblastic leukaemia (pre-B ALL) from healthy tissues suggesting LGR5 protein expression may provide a molecular handle for prognosis and treatment. We have developed highly specific, high affinity antibodies to the extracellular domain of human LGR5 (-LGR5) that detect high LGR5 protein levels in colorectal cancer (CRC), hepatocellular carcinoma (HCC), and pre-B ALL. In contrast, there is low to undetectable levels of LGR5 protein in normal colon and rectal epithelia, liver, ovarian tissues, brain and immune cell types. LGR5 is rapidly internalised from the plasma membrane and trafficked to intracellular vesicular compartments including lysosomes. Treatment of high LGR5-expressing CRC and pre-B ALL cancer cell lines with an antibody-drug conjugate version of -LGR5 (-LGR5-ADC) lead to effective cell killing at nanomolar concentrations. Interventional treatment of pre-B ALL tumours with -LGR5-ADC in vivo led to rapid tumour attrition. We further demonstrated the therapeutic utility of humanised -LGR5 by using the corresponding scFv fragment for the generation of -LGR5 chimeric antigen receptors (CARs) and a Bispecific T cell Engager (BiTE). -LGR5-CAR-NK cells were effective at killing LGR5-expressing cells while -LGR5/-CD3 BiTEs induce T cell activation and killing of NALM6 cells by cytotoxic CD8+ T cells. Taken together, this study establishes -LGR5-based therapeutic modalities that effectively discriminate and target CRC, HCC and pre-B ALL tumour cells. One Sentence SummaryWe generated novel antibodies against the cancer cell marker LGR5, validated diagnostic use in prioritizing specific cancer types for targeting, and developed antibody-based therapeutics.

cell biology↗

bioRxiv2021

Non-canonical Hedgehog signaling through L-type voltage gated Ca2+ channels controls CD8+ T cell killing

Cytotoxic CD8+ T lymphocytes (CTLs) are critical to the immune response against intracellular pathogens and cancer and act by eliminating infected and malignant cells through targeted secretion of cytotoxic granules. Hedgehog (Hh) signaling has been shown to be critical for CTL killing. Interestingly, Hh signaling in CD8+ T cells is not induced by extracellular Hh ligands but is initiated upon T cell receptor (TCR) engagement. How the TCR induces the Hh pathway independently of extracellular Hh ligands is unknown. Here we show that the Hh transcription factor Gli1 is essential for efficient CTL function and is induced downstream of the TCR by an extracellular Ca2+ influx selectively controlled by L-type voltage gated Ca2+ channels localized at the plasma membrane. We demonstrate that this novel mode of Hh signaling induction is independent of the canonical Hh pathway and represents the primary mechanism of Gli1 induction in naive CD8+ T cells, while CTLs can also activate Gli1 via MAP Kinase signaling. Importantly, we show that this L-type voltage gated Ca2+ channel-controlled Gli1 induction is functionally required for CTL killing in mice and humans. Gli inhibitors are currently in clinical trials against various cancers and our observations indicate that they likely inhibit the anti-tumor response. Significance statementCytotoxic CD8+ T cells (CTLs) kill infected and malignant cells by targeted secretion of cytotoxic granules. Hedgehog signaling is critical for effective CTL killing and is activated by the T cell receptor (TCR) independently of exogenous Hedgehog ligands. This study shows that Hedgehog transcription factor Gli1 is required for CTL killing and identifies L-type voltage gated Ca2+ channels (Cav1) as essential regulators of CTL killing in mouse and human, by virtue of their ability to activate Gli1 downstream of the TCR. This Cav1-Gli1 axis operates independently of canonical Hedgehog signaling. Our work suggests that caution is required when using Gli inhibitors, currently in trials as anti-cancer therapeutics, since they may dampen the anti-tumor response.

immunology↗

bioRxiv2020

Mutational inactivation of Apc in the intestinal epithelia compromises cellular organisation

The tumour suppressor adenomatous polyposis coli (Apc) regulates diverse effector pathways essential for cellular homeostasis. Truncating mutations in Apc, leading to the loss of its Wnt pathway and microtubule regulatory domains, are oncogenic in human and murine intestinal epithelia and drive malignant transformation. Whereas uncontrolled proliferation via Wnt pathway deregulation is an unequivocal consequence of oncogenic Apc mutations, it is not known whether loss of its other control systems contribute to tumorigenesis. Here we employ in vitro models of tumorigenesis to unmask the molecular barriers erected by Apc that maintain normal epithelial homeostasis in the murine intestinal epithelia. We determine that (i) enterocyte proliferation, (ii) microtubule dynamics and (iii) epithelial morphology are controlled by three independent molecular pathways, each corrupted by oncogenic Apc mutations. The key result of the study is to establish that Apc regulates three individual biological fates in the intestinal epithelia, through three distinct effector pathways, a significant advance to our understanding of normal tissue homeostasis, the molecular architecture of epithelial tissue and the aetiology of intestinal cancer.

cell biology↗

bioRxiv2018

Altered composition of the γδ T cell pool in lymph nodes during ageing enhances tumour growth

How age-associated decline of immune function leads to increased cancer incidence is poorly understood. Here, we have characterized the cellular composition of the{gamma}{delta} T cell pool in peripheral lymph nodes (pLNs) upon ageing. We found that ageing has minimal cell-intrinsic effects on function and global gene expression of{gamma}{delta} T cells, and TCR{gamma}{delta} diversity remained stable. However, ageing altered TCR{delta} chain usage and clonal structure of{gamma}{delta} T cell subsets. Importantly, IL-17-producing{gamma}{delta} 17 T cells dominated the{gamma}{delta} T cell pool of aged mice - mainly due to the selective expansion of V{gamma}6+{gamma}{delta} 17 T cells and augmented{gamma}{delta} 17-polarisation of V{gamma}4+ T cells. Expansion of the{gamma}{delta} 17 T cell compartment was supported by increased Interleukin-7 expression in the T cell zone of old mice. In a Lewis lung cancer model, pro-tumourigenic V{gamma}6+{gamma}{delta}17 T cells were exclusively activated in the tumour-draining LN and their infiltration into the tumour correlated with increased tumour size in aged mice. Thus, upon ageing, substantial compositional changes of{gamma}{delta} T cell pool in a dysregulated pLN microenvironment promote tumour growth.

immunology↗

bioRxiv2017

Primary cilia mediate diverse kinase inhibitor resistance mechanisms in cancer

Primary cilia are microtubule-based organelles that detect mechanical and chemical stimuli. Although cilia house a number of oncogenic molecules (including Smoothened, KRAS, EGFR, and PDGFR), their precise role in cancer remains unclear. We have interrogated the role of cilia in acquired and de novo resistance to a variety of kinase inhibitors, and found that in several examples, resistant cells are distinctly characterized by an increase in the number and/or length of cilia with altered structural features. Changes in cilia length seem to be linked to the lack of recruitment of Kif7 and IFT81 to cilia tips, and result in enhanced hedgehog pathway activation. Notably, Kif7 knockdown is sufficient to confer drug resistance in drug sensitive cells. Conversely, targeting of cilia length or integrity through genetic and pharmacological approaches overcomes kinase inhibitor resistance. The identification of a broad mechanism of pathway-unbiased drug resistance, represents a major advancement in oncology, and helps define a specific and important role for cilia in human cancer.

cell biology↗