DSpace Collection:
https://hdl.handle.net/10171/52057
2024-03-29T02:07:21ZThree-dimensional colon cancer organoids model the response to CEA-CD3 T-cell engagers
https://hdl.handle.net/10171/69001
Title: Three-dimensional colon cancer organoids model the response to CEA-CD3 T-cell engagers
Abstract: Rationale: The CEA-CD3 T cell bispecific antibody cibisatamab (CEA-TCB) is currently undergoing clinical trials. Here we study its performance against three-dimensional tumor organoids in cocultures with T cells as compared to a higher affinity CEACAM5-CD3 (CEACAM5-TCB) bispecific antibody using time-lapse confocal microscopy.
Methods: Pre-labelled spheroids derived from colon cancer cell lines and primary organoids derived from four colorectal cancer surgical specimens, which expressed different graded levels of CEA, were exposed in cocultures to T lymphocytes. Cocultures were treated with CEA-CD3 T-cell engagers and were followed by live confocal microscopy. Caspase 3 activation detected in real-time was used as an indicator of tumor cell death. Co-cultures were also set up with autologous tumor-associated fibroblasts to test the co-stimulatory effect of a fibroblast activated protein (FAP)- targeted 4-1BBL bispecific antibody fusion protein currently undergoing clinical trials.
Results: Tumor-cell killing of 3D colon carcinoma cultures was dependent on the levels of surface CEA expression, in such a way that the lower affinity agent (CEA-TCB) did not mediate killing by human preactivated T cells below a certain CEA expression threshold, while the high affinity construct (CEACAM5-TCB) remained active on the low CEA expressing organoids. Modelling heterogeneity in the levels of CEA expression by coculturing CEA high and low organoids showed measurable but weak bystander killing. Cocultures of tumor organoids, autologous fibroblasts and T cells allowed to observe a costimulatory effect of anti-FAP-4-1BBL both to release IFNγ and to attain more efficacious tumor cell killing.
Conclusion: Three-dimensional tumor cocultures with T cells using live confocal microscopy provide suitable models to test the requirements for colon-cancer redirected killing as elicited by CEA-targeted T-cell engagers undergoing clinical trials and treatment allow combinations to be tested in a relevant preclinical system.2022-01-01T00:00:00ZIntratumoral co-injection of the poly I:C-derivative BO-112 and a STING agonist synergize to achieve local and distant anti-tumor efficacy
https://hdl.handle.net/10171/68974
Title: Intratumoral co-injection of the poly I:C-derivative BO-112 and a STING agonist synergize to achieve local and distant anti-tumor efficacy
Abstract: Background BO-112 is a nanoplexed form of polyinosinic:polycytidylic acid that acting on toll-like receptor 3 (TLR3), melanoma differentiation-associated protein 5 (MDA5) and protein kinase RNA-activated (PKR) elicits rejection of directly injected transplanted tumors, but has only modest efficacy against distant untreated tumors. Its clinical activity has also been documented in early phase clinical trials. The 5,6-dimethylxanthenone-4-acetic acid (DMXAA) stimulator of interferon genes (STING) agonist shows a comparable pattern of efficacy when used via intratumoral injections.
Methods Mice subcutaneously engrafted with bilateral MC38 and B16.OVA-derived tumors were treated with proinflammatory immunotherapy agents known to be active when intratumorally delivered. The combination of BO-112 and DMXAA was chosen given its excellent efficacy and the requirements for antitumor effects were studied on selective depletion of immune cell types and in gene-modified mouse strains lacking basic leucine zipper ATF-like transcription factor 3 (BATF3), interferon-α/β receptor (IFNAR) or STING. Spatial requirements for the injections were studied in mice bearing three tumor lesions.
Results BO-112 and DMXAA when co-injected in one of the lesions of mice bearing concomitant bilateral tumors frequently achieved complete local and distant antitumor efficacy. Synergistic effects were contingent on CD8 T cell lymphocytes and dependent on conventional type 1 dendritic cells, responsiveness to type I interferon (IFN) and STING function in the tumor-bearing host. Efficacy was preserved even if BO-112 and DMXAA were injected in separate lesions in a manner able to control another untreated third-party tumor. Efficacy could be further enhanced on concurrent PD-1 blockade.
Conclusion Clinically feasible co-injections of BO-112 and a STING agonist attain synergistic efficacy able to eradicate distant untreated tumor lesions.2021-01-01T00:00:00ZThe involvement of ADAM10 in acantholysis in mucocutaneous pemphigus vulgaris depends on the autoantibody profile of each patient
https://hdl.handle.net/10171/68840
Title: The involvement of ADAM10 in acantholysis in mucocutaneous pemphigus vulgaris depends on the autoantibody profile of each patient
Abstract: Background
Acantholysis in pemphigus vulgaris (PV) may be triggered by desmoglein (Dsg) and non‐Dsg autoantibodies. The autoantibody profile of each patient results in distinct intracellular signalling patterns.
Objectives
Based on our previous findings, we aimed to elucidate whether PV acantholysis in a mouse model may be mediated by activation of a disintegrin and metalloproteinase 10 (ADAM10).
Methods
We used three PV‐IgG fractions from different patients containing high or low levels of anti‐Dsg1 and anti‐Dsg3 antibodies, and the presence or not of anti‐desmocollin (Dsc) antibodies, using a passive transfer mouse model of PV.
Results
Although all of the PV‐IgG fractions produced suprabasal acantholysis, only those containing anti‐Dsg1/3, but not anti‐Dsc2/3 antibodies, induced ADAM10 activation in a Src‐dependent way, and an increase in the epidermal growth factor (EGF) receptor ligands EGF and betacellulin (BTC). In contrast, the presence of anti‐Dsc2/3 antibodies, in addition to anti‐Dsg1/3, triggered earlier and ADAM10‐independent epidermal detachment, with no increase in EGF and BTC, which was associated with an earlier and more intense acantholysis.
Conclusions
All PV‐IgG fractions produced suprabasal acantholysis, but our results reveal that depending on the levels of anti‐Dsg antibodies or the presence of non‐Dsg antibodies, such as anti‐Dsc, more severe cell–cell epidermal detachment will occur at different times, and in an ADAM10‐dependent manner or not. Acantholysis in these different groups of patients with PV may be a consequence of the activation of specific intracellular mechanisms downstream of Autoantibodies binding to Dsg or non‐Dsg proteins, and therefore more specific therapeutic approaches in PV should be used.2020-01-01T00:00:00ZSerum levels of S-100 protein are directly proportional to the size, number, thickness and degree of cellularity of congenital melanocytic nevi
https://hdl.handle.net/10171/67902
Title: Serum levels of S-100 protein are directly proportional to the size, number, thickness and degree of cellularity of congenital melanocytic nevi
Abstract: To the Editor: Some patients with congenital melanocytic nevi (CMN) present progressive growth and
thickening, extracutaneous involvement (neurocutaneous melanocytosis, NCM) or neoplastic transformation (melanoma); and others remain stable or
even regress. There are no markers to assess progression or follow-up. Recently, we found S-100, a
protein which acts on cell differentiation and proliferation, elevated in CMN.1 S-100 is a ligand of the
RAGE pathway (related to the MAPK-pathway), and
low serum levels of soluble-RAGE were related to
poor survival in melanoma.2 Also SOX10, expressed
in melanocytes with high specificity, is useful in
detection, prognosis and treatment assessment of
melanoma.3 We explored if S-100, RAGE and SOX10
serum levels vary in children’s CMN and assessed
clinical or pathological correlations.2023-01-01T00:00:00Z