LIF regulates CXCL9 in tumor-associated macrophages and prevents CD8+ T cell tumor-infiltration impairing anti-PD1 therapy

Càncer; Macròfags associats al tumor: LIF; CD8Cáncer; Macrófagos asociados al tumor; CD8Cancer; Tumor-associated macrophages; CD8Cancer response to immunotherapy depends on the infiltration of CD8+ T cells and the presence of tumor-associated macrophages within tumors. Still, little is known about the determinants of these factors. We show that LIF assumes a crucial role in the regulation of CD8+ T cell tumor infiltration, while promoting the presence of protumoral tumor-associated macrophages. We observe that the blockade of LIF in tumors expressing high levels of LIF decreases CD206, CD163 and CCL2 and induces CXCL9 expression in tumor-associated macrophages. The blockade of LIF releases the epigenetic silencing of CXCL9 triggering CD8+ T cell tumor infiltration. The combination of LIF neutralizing antibodies with the inhibition of the PD1 immune checkpoint promotes tumor regression, immunological memory and an increase in overall survival

MCT4 blockade increases the efficacy of immune checkpoint blockade

Background & Aims Intratumoral lactate accumulation and acidosis impair T-cell function and antitumor immunity. Interestingly, expression of the lactate transporter monocarboxylate transporter (MCT) 4, but not MCT1, turned out to be prognostic for the survival of patients with rectal cancer, indicating that single MCT4 blockade might be a promising strategy to overcome glycolysis-related therapy resistance. Methods To determine whether blockade of MCT4 alone is sufficient to improve the efficacy of immune checkpoint blockade (ICB) therapy, we examined the effects of the selective MCT1 inhibitor AZD3965 and a novel MCT4 inhibitor in a colorectal carcinoma (CRC) tumor spheroid model co-cultured with blood leukocytes in vitro and the MC38 murine CRC model in vivo in combination with an antibody against programmed cell death ligand-1(PD-L1). Results Inhibition of MCT4 was sufficient to reduce lactate efflux in three-dimensional (3D) CRC spheroids but not in two-dimensional cell-cultures. Co-administration of the MCT4 inhibitor and ICB augmented immune cell infiltration, T-cell function and decreased CRC spheroid viability in a 3D co-culture model of human CRC spheroids with blood leukocytes. Accordingly, combination of MCT4 and ICB increased intratumoral pH, improved leukocyte infiltration and T-cell activation, delayed tumor growth, and prolonged survival in vivo. MCT1 inhibition exerted no further beneficial impact. Conclusions These findings demonstrate that single MCT4 inhibition represents a novel therapeutic approach to reverse lactic-acid driven immunosuppression and might be suitable to improve ICB efficacy

Molecular mechanisms involved in the immunomodulation induced by LIF in cancer

[eng] LIF is a pleiotropic cytokine that can act as an immunomodulatory factor in different biological processes, such as embryo implantation, organ transplantation tolerance and multiple sclerosis. Here, we have observed that in tumors expressing LIF, its blockade inhibits tumor growth, and that the anti-tumor effect of LIF neutralization is mediated by the polarization of tumor-associated myeloid cells (TAMCs). LIF sustains the expression of CCL22, MRC1 and CD163 in human healthy monocytes and in TAMCs from mouse syngeneic (non-small cell lung cancer, ovarian cancer and colorectal cancer) models, patient-derived xenograft models and organotypic cultures from glioblastoma. We show that LIF blockade down-regulates CCL22 secretion by TAMCs preventing regulatory T cell and inducing effector T and NK cell tumor infiltration, leading to an increase in tumor cell apoptosis and an anti-tumor response. A positive correlation between M2-like markers and LIF expression has been observed in glioblastoma patients, and high levels of these factors confer poor prognosis. Moreover, we have studied one of the multiple mechanisms through which LIF could modulate the inhibition of effector T and NK cells: the regulation of PD-1/PDL1 axis. We have seen that after anti-LIF treatment of tumor-bearing mice, the expression of PDL1 decreases in tumor cells and in TAMCs. Furthermore, a positive correlation between PDL1 and LIF expression has been observed in glioblastoma patients, and both proteins also correlate with CD44 expression, a marker for cancer-initiating cell (CIC) population. High levels of these three factors confer poor prognosis. Thus, we have found that LIF acts as an immune-suppressor in cancer recapitulating its normal function in other biological processes. LIF is a bridge between the innate and adaptive immune responses through the induction of CCL22 in TAMCs and subsequent regulation of regulatory and effector T and NK cells. Our results identify LIF as a promising immuno-oncology therapeutic target and establish the translational potential of anti-LIF agents.[cat] LIF és una citoquina que actua com un factor immunomodulador en diferents processos biologics, com en la implantacio de l'embrió, en la tolerancia en trasplantaments d'argans i en I'esclerosi múltiple. Hem observat que aquells tumors que expressen LIF, sofreixen una inhibició del creixement tumoral en resposta al seu bloqueig. Aquest efecte esta mitjangat per la polaritzacio de les cel•ules mieloides associades al tumor (TAMCs) cap a un fenotip M2. LIF sustenta l'expressio de CCL22, MRC1 i CD163 en monacits alats de sang periferica de donants sans, en TAMCs de models singenics de ratolf (cancer de pulmo, ovari i colon), en models xenagrafts derivats de pacients i en cultius organotipics de glioblastoma. Demostrem que el bloqueig de LIF disminueix la secrecio de CCL22 de les TAMCs, prevenint la infiltracio tumoral de les cel•ules T reguladores (Tregs) i induint el reclutament de cel•ules T efectores (Teff) i de natural killer (NKs), el que comporta un augment en I'apoptosi de les cel•ules tumorals. Hem observat una correlacio positiva entre l'expressio de marcadors del fenotip M2 i LIF en pacients de glioblastoma. Alts nivells d'aquests factors confereixen un mal pronastic. D'altra banda, hem estudiat un dels mecanismes mitjangant el qual LIF podria modular la inhibicio de les Teff i NKs: la regulacio PDL1. Hem observat que despres de tractar els ratolins amb l'anticas anti-LIF, l'expressio de PDL1 disminueix tant en les cel•ules tumorals corn en les TAMCs. A mes, hem trobat una correlacio positiva entre l'expressio de PDL1 i LIF en pacients amb glioblastoma, i ambdues protenes correlacionen amb CD44, un marcador d'una poblacio enriquida amb cel•ules iniciadores de tumor (CICs). Alts nivells d'aquests tres factors confereixen un mal pronastic. Aixf doncs, hem demostrat que LIF actua corn un supressor immunologic en processos tumorals, recapitulant la seva funcio normal en altres processos biologics. LIF es un pont entre la resposta immunolagica innata i I'adaptativa a traves de la induccio de la secrecio de CCL22 de les TAMCs, i la posterior regulacio de les Tregs, Teff i NKs. Els nostres resultats identifiquen LIF corn una diana terapeutica immuno-oncolagica prometedora i estableixen el potencial translacional d'agents inhibidors de LIF

Molecular mechanisms involved in the immunomodulation induced by LIF in cancer

[eng] LIF is a pleiotropic cytokine that can act as an immunomodulatory factor in different biological processes, such as embryo implantation, organ transplantation tolerance and multiple sclerosis. Here, we have observed that in tumors expressing LIF, its blockade inhibits tumor growth, and that the anti-tumor effect of LIF neutralization is mediated by the polarization of tumor-associated myeloid cells (TAMCs). LIF sustains the expression of CCL22, MRC1 and CD163 in human healthy monocytes and in TAMCs from mouse syngeneic (non-small cell lung cancer, ovarian cancer and colorectal cancer) models, patient-derived xenograft models and organotypic cultures from glioblastoma. We show that LIF blockade down-regulates CCL22 secretion by TAMCs preventing regulatory T cell and inducing effector T and NK cell tumor infiltration, leading to an increase in tumor cell apoptosis and an anti-tumor response. A positive correlation between M2-like markers and LIF expression has been observed in glioblastoma patients, and high levels of these factors confer poor prognosis. Moreover, we have studied one of the multiple mechanisms through which LIF could modulate the inhibition of effector T and NK cells: the regulation of PD-1/PDL1 axis. We have seen that after anti-LIF treatment of tumor-bearing mice, the expression of PDL1 decreases in tumor cells and in TAMCs. Furthermore, a positive correlation between PDL1 and LIF expression has been observed in glioblastoma patients, and both proteins also correlate with CD44 expression, a marker for cancer-initiating cell (CIC) population. High levels of these three factors confer poor prognosis. Thus, we have found that LIF acts as an immune-suppressor in cancer recapitulating its normal function in other biological processes. LIF is a bridge between the innate and adaptive immune responses through the induction of CCL22 in TAMCs and subsequent regulation of regulatory and effector T and NK cells. Our results identify LIF as a promising immuno-oncology therapeutic target and establish the translational potential of anti-LIF agents.[cat] LIF és una citoquina que actua com un factor immunomodulador en diferents processos biologics, com en la implantacio de l'embrió, en la tolerancia en trasplantaments d'argans i en I'esclerosi múltiple. Hem observat que aquells tumors que expressen LIF, sofreixen una inhibició del creixement tumoral en resposta al seu bloqueig. Aquest efecte esta mitjangat per la polaritzacio de les cel•ules mieloides associades al tumor (TAMCs) cap a un fenotip M2. LIF sustenta l'expressio de CCL22, MRC1 i CD163 en monacits alats de sang periferica de donants sans, en TAMCs de models singenics de ratolf (cancer de pulmo, ovari i colon), en models xenagrafts derivats de pacients i en cultius organotipics de glioblastoma. Demostrem que el bloqueig de LIF disminueix la secrecio de CCL22 de les TAMCs, prevenint la infiltracio tumoral de les cel•ules T reguladores (Tregs) i induint el reclutament de cel•ules T efectores (Teff) i de natural killer (NKs), el que comporta un augment en I'apoptosi de les cel•ules tumorals. Hem observat una correlacio positiva entre l'expressio de marcadors del fenotip M2 i LIF en pacients de glioblastoma. Alts nivells d'aquests factors confereixen un mal pronastic. D'altra banda, hem estudiat un dels mecanismes mitjangant el qual LIF podria modular la inhibicio de les Teff i NKs: la regulacio PDL1. Hem observat que despres de tractar els ratolins amb l'anticas anti-LIF, l'expressio de PDL1 disminueix tant en les cel•ules tumorals corn en les TAMCs. A mes, hem trobat una correlacio positiva entre l'expressio de PDL1 i LIF en pacients amb glioblastoma, i ambdues protenes correlacionen amb CD44, un marcador d'una poblacio enriquida amb cel•ules iniciadores de tumor (CICs). Alts nivells d'aquests tres factors confereixen un mal pronastic. Aixf doncs, hem demostrat que LIF actua corn un supressor immunologic en processos tumorals, recapitulant la seva funcio normal en altres processos biologics. LIF es un pont entre la resposta immunolagica innata i I'adaptativa a traves de la induccio de la secrecio de CCL22 de les TAMCs, i la posterior regulacio de les Tregs, Teff i NKs. Els nostres resultats identifiquen LIF corn una diana terapeutica immuno-oncolagica prometedora i estableixen el potencial translacional d'agents inhibidors de LIF

LIF regulates CXCL9 in tumor-associated macrophages and prevents CD8+ T cell tumor-infiltration impairing anti-PD1 therapy

Cancer response to immunotherapy depends on the infiltration of CD8 T cells and the presence of tumor-associated macrophages within tumors. Still, little is known about the determinants of these factors. We show that LIF assumes a crucial role in the regulation of CD8 T cell tumor infiltration, while promoting the presence of protumoral tumor-associated macrophages. We observe that the blockade of LIF in tumors expressing high levels of LIF decreases CD206, CD163 and CCL2 and induces CXCL9 expression in tumor-associated macrophages. The blockade of LIF releases the epigenetic silencing of CXCL9 triggering CD8 T cell tumor infiltration. The combination of LIF neutralizing antibodies with the inhibition of the PD1 immune checkpoint promotes tumor regression, immunological memory and an increase in overall survival

MCT4 blockade increases the efficacy of immune checkpoint blockade

Background & Aims Intratumoral lactate accumulation and acidosis impair T-cell function and antitumor immunity. Interestingly, expression of the lactate transporter monocarboxylate transporter (MCT) 4, but not MCT1, turned out to be prognostic for the survival of patients with rectal cancer, indicating that single MCT4 blockade might be a promising strategy to overcome glycolysis-related therapy resistance.Methods To determine whether blockade of MCT4 alone is sufficient to improve the efficacy of immune checkpoint blockade (ICB) therapy, we examined the effects of the selective MCT1 inhibitor AZD3965 and a novel MCT4 inhibitor in a colorectal carcinoma (CRC) tumor spheroid model co-cultured with blood leukocytes in vitro and the MC38 murine CRC model in vivo in combination with an antibody against programmed cell death ligand-1(PD-L1).Results Inhibition of MCT4 was sufficient to reduce lactate efflux in three-dimensional (3D) CRC spheroids but not in two-dimensional cell-cultures. Co-administration of the MCT4 inhibitor and ICB augmented immune cell infiltration, T-cell function and decreased CRC spheroid viability in a 3D co-culture model of human CRC spheroids with blood leukocytes. Accordingly, combination of MCT4 and ICB increased intratumoral pH, improved leukocyte infiltration and T-cell activation, delayed tumor growth, and prolonged survival in vivo. MCT1 inhibition exerted no further beneficial impact.Conclusions These findings demonstrate that single MCT4 inhibition represents a novel therapeutic approach to reverse lactic-acid driven immunosuppression and might be suitable to improve ICB efficacy

LIF regulates CXCL9 in tumor-associated macrophages and prevents CD8+ T cell tumor-infiltration impairing anti-PD1 therapy

Càncer; Macròfags associats al tumor: LIF; CD8Cáncer; Macrófagos asociados al tumor; CD8Cancer; Tumor-associated macrophages; CD8Cancer response to immunotherapy depends on the infiltration of CD8+ T cells and the presence of tumor-associated macrophages within tumors. Still, little is known about the determinants of these factors. We show that LIF assumes a crucial role in the regulation of CD8+ T cell tumor infiltration, while promoting the presence of protumoral tumor-associated macrophages. We observe that the blockade of LIF in tumors expressing high levels of LIF decreases CD206, CD163 and CCL2 and induces CXCL9 expression in tumor-associated macrophages. The blockade of LIF releases the epigenetic silencing of CXCL9 triggering CD8+ T cell tumor infiltration. The combination of LIF neutralizing antibodies with the inhibition of the PD1 immune checkpoint promotes tumor regression, immunological memory and an increase in overall survival

Discovery of Cycloalkyl[<i>c</i>]thiophenes as Novel Scaffolds for Hypoxia-Inducible Factor-2α Inhibitors

Hypoxia-inducible factors (HIFs) are heterodimeric transcription factors induced in diverse pathophysiological settings. Inhibition of HIF-2α has become a strategy for cancer treatment since the discovery that small molecules, upon binding into a small cavity of the HIF-2α PAS B domain, can alter its conformation and disturb the activity of the HIF dimer complex. Herein, the design, synthesis, and systematic SAR exploration of cycloalkyl[c]thiophenes as novel HIF-2α inhibitors are described, providing the first chemotype featuring an alkoxy–aryl scaffold. X-ray data confirmed the ability of these inhibitors to induce perturbation of key amino acids by appropriately presenting key pharmacophoric elements in the hydrophobic cavity. Selected compounds showed inhibition of VEGF-A secretion in cancer cells and prevention of Arg1 expression and activity in IL4-stimulated macrophages. Moreover, in vivo target gene modulation was demonstrated with compound 35r. Thus, the disclosed HIF-2α inhibitors represent valuable tools for investigating selective HIF-2α inhibition and its effect on tumor biology