A tumor-associated heparan sulfate-related glycosaminoglycan promotes the generation of functional regulatory T cells

14 p.-8 fig.Functional Tregs play a key role in tumor development and progression, representing a major barrier to anticancer immunity. The mechanisms by which Tregs are generated in cancer and the influence of the tumor microenvironment on these processes remain incompletely understood. Herein, by using NMR, chemoenzymatic structural assays and a plethora of in vitro and in vivo functional analyses, we demonstrate that the tumoral carbohydrate A10 (Ca10), a cell-surface carbohydrate derived from Ehrlich’s tumor (ET) cells, is a heparan sulfate-related proteoglycan that enhances glycolysis and promotes the development of tolerogenic features in human DCs. Ca10-stimulated human DCs generate highly suppressive Tregs by mechanisms partially dependent on metabolic reprogramming, PD-L1, IL-10, and IDO. Ca10 also reprograms the differentiation of human monocytes into DCs with tolerogenic features. In solid ET-bearing mice, we found positive correlations between Ca10 serum levels, tumor size and splenic Treg numbers. Administration of isolated Ca10 also increases the proportion of splenic Tregs in tumor-free mice. Remarkably, we provide evidence supporting the presence of a circulating human Ca10 counterpart (Ca10H) and show, for the first time, that serum levels of Ca10H are increased in patients suffering from different cancer types compared to healthy individuals. Of note, these levels are higher in prostate cancer patients with bone metastases than in prostate cancer patients without metastases. Collectively, we reveal novel molecular mechanisms by which heparan sulfate-related structures associated with tumor cells promote the generation of functional Tregs in cancer. The discovery of this novel structural-functional relationship may open new avenues of research with important clinical implications in cancer treatment.The authors’ laboratories were supported by grants FEI 16_60 from Art.83 4154605 (138/2012)(24/2013), SAF-2017-84978-R from MINECO (Spain) and PID2020-114396RB-I00 from Ministerio de Ciencia e Innovación (Spain) to OP and PID2021-123781OB-C22 to FJC funded by MCIN/AEI/10.13039/501100011033 (Spain) and RTC-2015-3805-1 from MINECO to Inmunotek SL.Peer reviewe

Estudio de la respuesta inmunitaria frente al complejo silicio mesoporoso-Ca10 (MSMPs-Ca10) como vacuna antineoplásica

Tesis inédita de la Universidad Complutense de Madrid, Facultad de Medicina, leída el 21-12-2022En las últimas décadas, la inmunoterapia ha irrumpido con fuerza como alternativa para tratar el cáncer, superando las limitaciones de los tratamientos convencionales. Este tipo de terapia es capaz de modular el potencial del sistema inmunitario para la inducción de respuestas antitumorales. Un tipo de inmunoterapia son las vacunas antineoplásicas, las cuales pueden ayudar a tratar o prevenir ciertos tipos de cáncer. Durante la tumorogénesis, las células pueden sufrir modificaciones en los proteoglicanos presentes en el glicocálix mediante procesos de glicosilación incompleta o aberrante, dando lugar a nuevos antígenos potencialmente reconocibles por el sistema inmunitario. Este tipo de antígenos tumorales juegan un papel importante en la metástasis de células tumorales y la transducción de señales, y la sobreexpresión de éstos en la superficie celular se correlaciona frecuentemente con mal pronóstico. Un ejemplo de ello es el antígeno Ca10, una macromolécula de alto peso molecular caracterizada como un proteoglicano de heparán sulfato que contiene el epítopo de naturaleza sacarídica definido por el anticuerpo monoclonal A10. Este epítopo de tipo carbohidrato se encuentra expresado en la superficie de membrana de células del tumor murino de Ehrlich (ET) y en algunos tumores epiteliales humanos de origen glandular. Además, también se detecta en el suero de determinados pacientes con estos tipos de tumores. Por todo ello, el Ca10 es una potencial diana para el desarrollo de una vacuna antitumoral. Para su desarrollo, el Ca10 fue formulado con un potencial nuevo adyuvante basado en micropartículas de silicio mesoporoso (MSMPs-Ca10), demostrando inducir una eficiente respuesta humoral y de resistencia tumoral. Las micropartículas mesoporosas de silicio (MSMPs) ofrecen grandes ventajas como adyuvantes: poseen un tamaño de poro ajustable y una naturaleza biocompatible y biodegradable. Sabemos del gran potencial de estos materiales para estimular células presentadoras de antígeno (APCs), demostrando mejorar la capacidad de presentación antigénica específica de células dendríticas (DCs) humanas a células T CD8. Las DCs están involucradas en la consecución de respuestas innatas y adaptativas, pudiendo facilitar respuestas específicas de antígeno por parte de células T, en el caso de respuestas T-dependientes. La respuesta humoral frente al Ca10, debido a su mayoritaria estructura sacarídica con epítopos repetitivos, se ha adscrito como T-independiente. El reconocimiento de antígenos T-independientes de tipo II depende en mayor medida de Linfocitos B de la zona marginal del bazo y linfocitos B1...In recent decades, immunotherapy has strongly emerged as an alternative for treating cancer, overcoming the limitations of conventional treatments. This type of therapy can modulate the potential of the immune system to induce antitumor responses. One type of immunotherapy is antineoplastic vaccines, which can help treat or prevent certain types of cancer.During tumorogenesis, cells can undergo modifications in the proteoglycans present in the glycocalyx, through incomplete or aberrant glycosylation processes, giving rise to new antigens potentially recognizable by the immune system. These types of tumor antigens play an important role in tumor cell metastasis and signal transduction, and their overexpression on the cell surface frequently correlates with poor prognosis. An example of this is the Ca10 antigen, a high molecular weight macromolecule characterized as a heparan sulfate proteoglycan containing the saccharide epitope defined by the monoclonal antibody A10. This carbohydrate-type epitope is expressed on the membrane surface of murine Ehrlich tumor (ET) cells and in some human epithelial tumors of glandular origin. In addition, it is also detected in the serum of certain patients with these types of tumors. Therefore, Ca10 is a potential target for the development of an anti-tumor vaccine. For its development, Ca10 was formulated with a potential new adjuvant based on mesoporous silica microparticles (MSMPs-Ca10), showing to induce an efficient humoral response and tumor resistance. Mesoporous silica microparticles (MSMPs) offer great advantages as adjuvants: MSMPs possess an adjustable pore size and a biocompatible and biodegradable nature. We know of the great potential of these materials to stimulate antigen presenting cells (APCs), demonstrating to enhance the specific antigenic presentation capacity of human dendritic cells (DCs) to CD8 T cells. DCs are involved in eliciting innate and adaptive responses and can facilitate antigen-specific responses by T cells in the case of T-dependent responses. The humoral response to Ca10, due to its mostly saccharide structure with repetitive epitopes, has been ascribed as T-independent. The recognition of type II T-independent antigens depends to a greater extent on B lymphocytes of the marginal zone of the spleen and B1 lymphocytes...Fac. de MedicinaTRUEunpu

Monocyte-Derived Dendritic Cells Differentiated in the Presence of Lenalidomide Display a Semi-Mature Phenotype, Enhanced Phagocytic Capacity, and Th1 Polarization Capability

Lenalidomide is an analog of thalidomide, with potent anticancer activity demonstrated in several hematological malignancies. It has immunomodulatory properties, being able to enhance the activation of different types of immune cells, which results in antitumor activities. Dendritic cells (DCs) are pivotal in the immune response, and different immunotherapeutic approaches targeting these cells are being developed. Since little is known about the effect of lenalidomide on DCs, the goal of the present work was to investigate the phenotype and function of human monocyte-derived DCs differentiated in the presence of lenalidomide (L-DCs). Our results showed that L-DCs display a unique phenotype, with increased cell surface expression of some maturation markers such as CD1d, CD83, CD86, and HLA-DR. This phenotype correlates with a lower expression of the E3 ubiquitin-ligase MARCH-I in L-DCs, upregulating the cell surface expression of CD86 and HLA-DR. In addition, immature L-DCs express higher amounts of DC-SIGN on the cell surface than control immature DCs. After LPS stimulation, production of IL-6 and TNF-α was severely decreased, whereas IL-12 and IL-10 secretion was dramatically upregulated in L-DCs, compared to that in the controls. Functionally, L-DCs are more effectively recognized by NKT cells in cytotoxicity experiments. Furthermore, L-DCs display higher opsonin-independent antigen uptake capability than control DCs. Mixed lymphocyte reaction experiments showed that L-DCs could stimulate naïve CD4 T-cells, polarizing them toward a predominant Th1 phenotype. In summary, DCs derived from monocytes in the presence of lenalidomide present a semi-mature phenotype, increased phagocytic capacity, reduced production of proinflammatory cytokines, and the ability to polarize T-cells toward predominant Th1-type responses; these are qualities that might be useful in the development of new immunotherapeutic treatments

Monocyte-Derived Dendritic Cells Differentiated in the Presence of Lenalidomide Display a Semi-Mature Phenotype, Enhanced Phagocytic Capacity, and Th1 Polarization Capability

Lenalidomide is an analog of thalidomide, with potent anticancer activity demonstrated in several hematological malignancies. It has immunomodulatory properties, being able to enhance the activation of different types of immune cells, which results in antitumor activities. Dendritic cells (DCs) are pivotal in the immune response, and different immunotherapeutic approaches targeting these cells are being developed. Since little is known about the effect of lenalidomide on DCs, the goal of the present work was to investigate the phenotype and function of human monocyte-derived DCs differentiated in the presence of lenalidomide (L-DCs). Our results showed that L-DCs display a unique phenotype, with increased cell surface expression of some maturation markers such as CD1d, CD83, CD86, and HLA-DR. This phenotype correlates with a lower expression of the E3 ubiquitin-ligase MARCH-I in L-DCs, upregulating the cell surface expression of CD86 and HLA-DR. In addition, immature L-DCs express higher amounts of DC-SIGN on the cell surface than control immature DCs. After LPS stimulation, production of IL-6 and TNF-α was severely decreased, whereas IL-12 and IL-10 secretion was dramatically upregulated in L-DCs, compared to that in the controls. Functionally, L-DCs are more effectively recognized by NKT cells in cytotoxicity experiments. Furthermore, L-DCs display higher opsonin-independent antigen uptake capability than control DCs. Mixed lymphocyte reaction experiments showed that L-DCs could stimulate naïve CD4 T-cells, polarizing them toward a predominant Th1 phenotype. In summary, DCs derived from monocytes in the presence of lenalidomide present a semi-mature phenotype, increased phagocytic capacity, reduced production of proinflammatory cytokines, and the ability to polarize T-cells toward predominant Th1-type responses; these are qualities that might be useful in the development of new immunotherapeutic treatments.Ministerio de Economía y CompetitividadUniversidad Complutense de MadridDepto. de Inmunología, Oftalmología y ORLFac. de MedicinaTRUEpu

Image_1_Monocyte-Derived Dendritic Cells Differentiated in the Presence of Lenalidomide Display a Semi-Mature Phenotype, Enhanced Phagocytic Capacity, and Th1 Polarization Capability.PDF

<p>Lenalidomide is an analog of thalidomide, with potent anticancer activity demonstrated in several hematological malignancies. It has immunomodulatory properties, being able to enhance the activation of different types of immune cells, which results in antitumor activities. Dendritic cells (DCs) are pivotal in the immune response, and different immunotherapeutic approaches targeting these cells are being developed. Since little is known about the effect of lenalidomide on DCs, the goal of the present work was to investigate the phenotype and function of human monocyte-derived DCs differentiated in the presence of lenalidomide (L-DCs). Our results showed that L-DCs display a unique phenotype, with increased cell surface expression of some maturation markers such as CD1d, CD83, CD86, and HLA-DR. This phenotype correlates with a lower expression of the E3 ubiquitin-ligase MARCH-I in L-DCs, upregulating the cell surface expression of CD86 and HLA-DR. In addition, immature L-DCs express higher amounts of DC-SIGN on the cell surface than control immature DCs. After LPS stimulation, production of IL-6 and TNF-α was severely decreased, whereas IL-12 and IL-10 secretion was dramatically upregulated in L-DCs, compared to that in the controls. Functionally, L-DCs are more effectively recognized by NKT cells in cytotoxicity experiments. Furthermore, L-DCs display higher opsonin-independent antigen uptake capability than control DCs. Mixed lymphocyte reaction experiments showed that L-DCs could stimulate naïve CD4 T-cells, polarizing them toward a predominant Th1 phenotype. In summary, DCs derived from monocytes in the presence of lenalidomide present a semi-mature phenotype, increased phagocytic capacity, reduced production of proinflammatory cytokines, and the ability to polarize T-cells toward predominant Th1-type responses; these are qualities that might be useful in the development of new immunotherapeutic treatments.</p

Transcriptional regulation by NR5A2 links differentiation and inflammation in the pancreas

Chronic inflammation increases the risk of developing one of several types of cancer. Inflammatory responses are currently thought to be controlled by mechanisms that rely on transcriptional networks that are distinct from those involved in cell differentiation1,2,3. The orphan nuclear receptor NR5A2 participates in a wide variety of processes, including cholesterol and glucose metabolism in the liver, resolution of endoplasmic reticulum stress, intestinal glucocorticoid production, pancreatic development and acinar differentiation4,5,6,7,8. In genome-wide association studies9,10, single nucleotide polymorphisms in the vicinity of NR5A2 have previously been associated with the risk of pancreatic adenocarcinoma. In mice, Nr5a2 heterozygosity sensitizes the pancreas to damage, impairs regeneration and cooperates with mutant Kras in tumour progression11. Here, using a global transcriptomic analysis, we describe an epithelial-cell-autonomous basal pre-inflammatory state in the pancreas of Nr5a2+/− mice that is reminiscent of the early stages of pancreatitis-induced inflammation and is conserved in histologically normal human pancreases with reduced expression of NR5A2 mRNA. In Nr5a2+/−mice, NR5A2 undergoes a marked transcriptional switch, relocating from differentiation-specific to inflammatory genes and thereby promoting gene transcription that is dependent on the AP-1 transcription factor. Pancreatic deletion of Jun rescues the pre-inflammatory phenotype, as well as binding of NR5A2 to inflammatory gene promoters and the defective regenerative response to damage. These findings support the notion that, in the pancreas, the transcriptional networks involved in differentiation-specific functions also suppress inflammatory programmes. Under conditions of genetic or environmental constraint, these networks can be subverted to foster inflammation

Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries

Background Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks. Methods The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned. Results A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P &lt; 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31). Conclusion Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)