The target antigen determines the mechanism of acquired resistance to T cell-based therapies

Cancer; Antigen; ResistanceCáncer; Antígeno; ResistenciaCàncer; Antigen; ResistènciaDespite the revolution of immunotherapy in cancer treatment, patients eventually progress due to the emergence of resistance. In this scenario, the selection of the tumor antigen can be decisive in the success of the clinical response. T cell bispecific antibodies (TCBs) are engineered molecules that include binding sites to the T cell receptor and to a tumor antigen. Using gastric CEA+/HER2+ MKN45 cells and TCBs directed against CEA or HER2, we show that the mechanism of resistance to a TCB is dependent on the tumor antigen. Acquired resistant models to a high-affinity-CEA-targeted TCB exhibit a reduction of CEA levels due to transcriptional silencing, which is reversible upon 5-AZA treatment. In contrast, a HER2-TCB resistant model maintains HER2 levels and exhibit a disruption of the interferon-gamma signaling. These results will help in the design of combinatorial strategies to increase the efficacy of cancer immunotherapies and to anticipate and overcome resistances.This work was supported by Asociación Española Contra el Cancer (AECC), Breast Cancer Research Foundation (BCRF-21-008), and Instituto de Salud Carlos III (PI19/01181). A.M.S. was funded by the Spanish Government (PFIS FI20/00188). B.M. was funded by a fellowship from PERIS (Departament de Salut, Generalitat de Catalunya). M.R.A. was funded by Agency for Management of University and Research Grants (AGAUR, 2022 FI_B2 00080). P.O.R. was funded by the BBVA. E.J.A. was funded by the AECC (POSTD211413AREN). VHIO acknowledges the Cellex Foundation for providing research facilities and equipment, the Centro de Investigación Biomédica en Red de Cáncer (CIBERONC) from the Institute of Health Carlos III (ISCIII), and the Department of Health (Generalitat de Catalunya, SLT008/18/00198 SLT008/18/00205) for their support on this research. The authors acknowledge financial support from the State Agency for Research (Agencia Estatal de Investigación) (CEX2020-001024-S/AEI/10.13039/501100011033) and for the Cancer Immunology and Immunotherapy (CAIMI-2) program funded by BBVA Foundation. We would like to remark the funding from B.M PERIS (Spain). The authors thank Dr. Anne Freimoser-Grundschober and Roche for helping provide the TCBs. The graphical abstract was created with BioRender.com

Assessing Blood-Based Biomarkers to Define a Therapeutic Window for Natalizumab

Natalizumab is a monoclonal antibody that binds CD49d. Although it is one of the most effective treatments for Relapsing-Remitting Multiple Sclerosis (RRMS), a dosing regimen has not been optimized for safety and efficacy in individual patients. We aimed to identify biomarkers to monitor Natalizumab treatment and to establish a personalized dose utilizing an ongoing longitudinal study in 29 RRMS patients under Natalizumab with standard interval dose (SD) of 300 mg/4 wks or extended interval dose (EID) of 300 mg/6 wks. Blood samples were analyzed by flow cytometry to determine CD49d saturation and expression in several T and B lymphocytes subpopulations. Each patient was analyzed at two different timepoints separated by 3 Natalizumab administrations. Natalizumab and sVCAM-1 levels in serum were also analyzed using ELISA. To determine the reproducibility of various markers, two different timepoints were compared and no significant differences were observed for CD49d expression nor for saturation; SD patients had higher saturation levels (~80%) than EID patients (~60%). A positive correlation exists between CD49d saturation and Natalizumab serum levels. CD49d expression and saturation are stable parameters that could be used as biomarkers in the immunomonitoring of Natalizumab treatment. Moreover, Natalizumab and sVCAM-1 serum levels could be used to optimize an individual's dosing schedule

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Redirection of T cells as immunotherapeutic approach against HER2-positive breast cancer

Tot hi els èxits de les teràpies contra HER2, l’aparició de resistències primàries i adquirides limita el potencial antitumoral dels tractaments actuals contra el càncer de mama HER2-positiu. A més a més, l’expressió de HER2 en teixits epitelials normals pot comportar l’aparició d’efectes secundaris post-tractament. Per tant, hi ha una necessitat de desenvolupar tractaments més segurs i eficaços contra tumors HER2-positius. La immunoteràpia constitueix un nou grup de teràpies que han revolucionat el camp del càncer. Una estratègia per activar el sistema immunitari contra tumors consisteix en la redirecció de cèl·lules T a través d’anticossos biespecífics o CAR Ts. Tot hi el potencial de les noves estratègies terapèutiques, fins ara, només s’han aprovat CAR Ts i TCBs per pacients amb malalties hematològiques. Un dels problemes en el desenvolupament de CAR Ts i TCBs dirigits a tumors sòlids és la falta d’antígens realment específics de tumor. Degut a aquesta falta, la majoria de CAR Ts i TCBs desenvolupats fins ara, van dirigits a antígens associats a tumor, els quals també es troben en teixits sà, encara que a nivells més baixos. Aquests CAR Ts i TCBs han causat greus efectes secundaris o inclús toxicitats letals degut a la falta d’especificitat tumoral. En aquesta tesis, demostrem que p95HER2, una forma truncada de HER2, és un antigen específic de tumor, expressat en un grup de pacients de càncer de mama HER2-positiu. A més a més, proposem p95HER2-TCB i p95HER2 CAR Ts com estratègies terapèutiques immunes que podrien suposar una opció més segura que les immunoteràpies que es dirigeixen a antígens associats a tumor.A pesar del éxito de las terapias contra HER2, la aparición de resistencias primarias y adquiridas limita el potencial antitumoral de los tratamientos actuales contra el cáncer de mama HER2-positivo. Además, la expresión de HER2 en tejidos epiteliales normales puede conllevar la aparición de efectos secundarios post-tratamiento. Por todo ello, hay una necesidad de desarrollar tratamientos más seguros y eficaces contra tumores HER2-positivos. La inmunoterapia constituye un nuevo grupo de terapias que han revolucionado el campo del cáncer. Una de las estrategias para activar el sistema inmunitario contra tumores consiste en la redirección de células T a través de anticuerpos biespecíficos o de CAR Ts. A pesar del potencial de estas nuevas estrategias terapéuticas, hasta ahora, solo se han aprobado CAR Ts y TCBs para pacientes con enfermedades hematológicas. Uno de los principales problemas en el desarrollo de CAR Ts y TCBs dirigidos a tumores sólidos es la falta de antígenos realmente específicos de tumor. Debido a esta falta, la mayoría de CAR Ts y TCBs desarrollados hasta ahora van dirigidos a antígenos asociados a tumor, los cuales también se encuentran en tejido sano, aunque a niveles mas bajos. Estos CAR Ts y TCBs han causado graves efectos secundarios o incluso toxicidades letales debido a la falta de especificidad de tejido tumoral. En esta tesis, demostramos que p95HER2, una forma truncada de HER2, es un antígeno específico de tumor, expresado en un grupo de pacientes de cáncer de mama HER2-positivo. Además, proponemos p95HER2-TCB y p95HER2 CAR Ts como estrategias terapéuticas inmunes que podrían suponer una opción más segura que las inmunoterapias que se dirigen a antígenos asociados a tumor.Despite the success of anti-HER2 therapies, the appearance of primary and acquired resistances is limiting the antitumor potential of the current treatments for HER2-positive breast cancer. Moreover, the expression of the tumour-associated antigen HER2 in normal epithelial tissues may cause undesired side effects upon anti-HER2 treatments. Therefore, there is a clinical need to develop more effective and safer treatments against HER2-driven tumours. Immunotherapy is one of the most promising field in cancer treatment. One of the strategies to activate the immune system against tumours is the redirection of T cells via bispecific antibodies or CAR T technology. Despite the potential of these therapeutic strategies, to date, only CAR Ts and TCBs targeting certain haematological malignancies have been approved to treat patients. One of the main hurdles in the development of CAR Ts and TCBs for solid tumours is the scarcity of tumour-specific antigens. Because of this shortage, CAR Ts and TCBs have been directed against tumour-associated antigens, which are also expressed in normal tissues, albeit at lower levels. These CAR Ts and TCBs have caused serious or even fatal toxicities because of their “on-target off-tumour” effects on normal tissues. In this thesis, we demonstrate that p95HER2, a truncated form of HER2, is a bona fide tumour-specific antigen, expressed by a subset of HER2-positive breast cancer patients. Moreover, we prove that the redirection of T lymphocytes via p95HER2-TCB or p95HER2 CAR Ts could be a safe therapeutic option against breast cancer, in contrast to other immune therapies targeting tumour-associated antigens

Redirection of T cells as immunotherapeutic approach against HER2-positive breast cancer /

Departament responsable de la tesi: Departament de Bioquímica i Biologia Molecular.Tot hi els èxits de les teràpies contra HER2, l'aparició de resistències primàries i adquirides limita el potencial antitumoral dels tractaments actuals contra el càncer de mama HER2-positiu. A més a més, l'expressió de HER2 en teixits epitelials normals pot comportar l'aparició d'efectes secundaris post-tractament. Per tant, hi ha una necessitat de desenvolupar tractaments més segurs i eficaços contra tumors HER2-positius. La immunoteràpia constitueix un nou grup de teràpies que han revolucionat el camp del càncer. Una estratègia per activar el sistema immunitari contra tumors consisteix en la redirecció de cèl·lules T a través d'anticossos biespecífics o CAR Ts. Tot hi el potencial de les noves estratègies terapèutiques, fins ara, només s'han aprovat CAR Ts i TCBs per pacients amb malalties hematològiques. Un dels problemes en el desenvolupament de CAR Ts i TCBs dirigits a tumors sòlids és la falta d'antígens realment específics de tumor. Degut a aquesta falta, la majoria de CAR Ts i TCBs desenvolupats fins ara, van dirigits a antígens associats a tumor, els quals també es troben en teixits sà, encara que a nivells més baixos. Aquests CAR Ts i TCBs han causat greus efectes secundaris o inclús toxicitats letals degut a la falta d'especificitat tumoral. En aquesta tesis, demostrem que p95HER2, una forma truncada de HER2, és un antigen específic de tumor, expressat en un grup de pacients de càncer de mama HER2-positiu. A més a més, proposem p95HER2-TCB i p95HER2 CAR Ts com estratègies terapèutiques immunes que podrien suposar una opció més segura que les immunoteràpies que es dirigeixen a antígens associats a tumor.A pesar del éxito de las terapias contra HER2, la aparición de resistencias primarias y adquiridas limita el potencial antitumoral de los tratamientos actuales contra el cáncer de mama HER2-positivo. Además, la expresión de HER2 en tejidos epiteliales normales puede conllevar la aparición de efectos secundarios post-tratamiento. Por todo ello, hay una necesidad de desarrollar tratamientos más seguros y eficaces contra tumores HER2-positivos. La inmunoterapia constituye un nuevo grupo de terapias que han revolucionado el campo del cáncer. Una de las estrategias para activar el sistema inmunitario contra tumores consiste en la redirección de células T a través de anticuerpos biespecíficos o de CAR Ts. A pesar del potencial de estas nuevas estrategias terapéuticas, hasta ahora, solo se han aprobado CAR Ts y TCBs para pacientes con enfermedades hematológicas. Uno de los principales problemas en el desarrollo de CAR Ts y TCBs dirigidos a tumores sólidos es la falta de antígenos realmente específicos de tumor. Debido a esta falta, la mayoría de CAR Ts y TCBs desarrollados hasta ahora van dirigidos a antígenos asociados a tumor, los cuales también se encuentran en tejido sano, aunque a niveles mas bajos. Estos CAR Ts y TCBs han causado graves efectos secundarios o incluso toxicidades letales debido a la falta de especificidad de tejido tumoral. En esta tesis, demostramos que p95HER2, una forma truncada de HER2, es un antígeno específico de tumor, expresado en un grupo de pacientes de cáncer de mama HER2-positivo. Además, proponemos p95HER2-TCB y p95HER2 CAR Ts como estrategias terapéuticas inmunes que podrían suponer una opción más segura que las inmunoterapias que se dirigen a antígenos asociados a tumor.Despite the success of anti-HER2 therapies, the appearance of primary and acquired resistances is limiting the antitumor potential of the current treatments for HER2-positive breast cancer. Moreover, the expression of the tumour-associated antigen HER2 in normal epithelial tissues may cause undesired side effects upon anti-HER2 treatments. Therefore, there is a clinical need to develop more effective and safer treatments against HER2-driven tumours. Immunotherapy is one of the most promising field in cancer treatment. One of the strategies to activate the immune system against tumours is the redirection of T cells via bispecific antibodies or CAR T technology. Despite the potential of these therapeutic strategies, to date, only CAR Ts and TCBs targeting certain haematological malignancies have been approved to treat patients. One of the main hurdles in the development of CAR Ts and TCBs for solid tumours is the scarcity of tumour-specific antigens. Because of this shortage, CAR Ts and TCBs have been directed against tumour-associated antigens, which are also expressed in normal tissues, albeit at lower levels. These CAR Ts and TCBs have caused serious or even fatal toxicities because of their “on-target off-tumour” effects on normal tissues. In this thesis, we demonstrate that p95HER2, a truncated form of HER2, is a bona fide tumour-specific antigen, expressed by a subset of HER2-positive breast cancer patients. Moreover, we prove that the redirection of T lymphocytes via p95HER2-TCB or p95HER2 CAR Ts could be a safe therapeutic option against breast cancer, in contrast to other immune therapies targeting tumour-associated antigens

The target antigen determines the mechanism of acquired resistance to T cell-based therapies

Despite the revolution of immunotherapy in cancer treatment, patients eventually progress due to the emergence of resistance. In this scenario, the selection of the tumor antigen can be decisive in the success of the clinical response. T cell bispecific antibodies (TCBs) are engineered molecules that include binding sites to the T cell receptor and to a tumor antigen. Using gastric CEA+/HER2+ MKN45 cells and TCBs directed against CEA or HER2, we show that the mechanism of resistance to a TCB is dependent on the tumor antigen. Acquired resistant models to a high-affinity-CEA-targeted TCB exhibit a reduction of CEA levels due to transcriptional silencing, which is reversible upon 5-AZA treatment. In contrast, a HER2-TCB resistant model maintains HER2 levels and exhibit a disruption of the interferon-gamma signaling. These results will help in the design of combinatorial strategies to increase the efficacy of cancer immunotherapies and to anticipate and overcome resistances

Acquired cancer cell resistance to T cell bispecific antibodies and CAR T targeting HER2 through JAK2 down-modulation

Immunotherapy has raised high expectations in the treatment of virtually every cancer. Many current efforts are focused on ensuring the efficient delivery of active cytotoxic cells to tumors. It is assumed that, once these active cytotoxic cells are correctly engaged to cancer cells, they will unfailingly eliminate the latter, provided that inhibitory factors are in check. T cell bispecific antibodies (TCBs) and chimeric antigen receptors (CARs) offer an opportunity to test this assumption. Using TCB and CARs directed against HER2, here we show that disruption of interferon-gamma signaling confers resistance to killing by active T lymphocytes. The kinase JAK2, which transduces the signal initiated by interferon-gamma, is a component repeatedly disrupted in several independently generated resistant models. Our results unveil a seemingly widespread strategy used by cancer cells to resist clearance by redirected lymphocytes. In addition, they open the possibility that long-term inhibition of interferon-gamma signaling may impair the elimination phase of immunoediting and, thus, promote tumor progression

LCOR mediates interferon-independent tumor immunogenicity and responsiveness to immune-checkpoint blockade in triple-negative breast cancer

Ligand-dependent corepressor (LCOR) mediates normal and malignant breast stem cell differentiation. Cancer stem cells (CSCs) generate phenotypic heterogeneity and drive therapy resistance, yet their role in immunotherapy is poorly understood. Here we show that immune-checkpoint blockade (ICB) therapy selects for LCOR CSCs with reduced antigen processing/presentation machinery (APM) driving immune escape and ICB resistance in triple-negative breast cancer (TNBC). We unveil an unexpected function of LCOR as a master transcriptional activator of APM genes binding to IFN-stimulated response elements (ISREs) in an IFN signaling-independent manner. Through genetic modification of LCOR expression, we demonstrate its central role in modulation of tumor immunogenicity and ICB responsiveness. In TNBC, LCOR associates with ICB clinical response. Importantly, extracellular vesicle (EV) Lcor-messenger RNA therapy in combination with anti-PD-L1 overcame resistance and eradicated breast cancer metastasis in preclinical models. Collectively, these data support LCOR as a promising target for enhancement of ICB efficacy in TNBC, by boosting of tumor APM independently of IFN

The Need for the Closer Monitoring of Novel Drugs in MS: A Siponimod Retrospective Cohort Study (Realhes Study)

Background: Severe cases of lymphopenia have been reported during siponimod clinical trials, which may negatively impact its benefit/risk profile. Objective: We aimed to evaluate the incidence of lymphopenia following the initiation of siponimod treatment in clinical practice. The secondary objectives included the analysis of factors predisposing to and the clinical relevance of lymphopenia events. Methods: In this multicenter retrospective cohort study, information collected from the medical records of 129 patients with MS from 15 tertiary hospitals in Spain who initiated treatment with Siponimod were followed-up for at least 3 months, including at least one lymphocyte count evaluation per patient. Results: Of the 129 patients, 121 (93.6%) reported lymphopenia events, including 110 (85.3%) with grade <= 3 and 11 (8.5%) with grade 4 lymphopenia, higher than those reported in the pivotal clinical trial (73.3% and 3.3% for grade <= 3 and grade 4 lymphopenia, respectively). The study included an unexpectedly high proportion of male subjects (72.9%), which might have led to an underestimation of the actual magnitude of the risk. Conclusions: In this study, the incidence and severity of lymphopenia after starting siponimod treatment were higher than those reported in previous clinical trials. Therefore, our results reinforce the need for the closer monitoring of novel MS drugs in clinical practice, as well as larger and longer follow-up studies to properly characterize this risk

Assessment of two complementary influenza surveillance systems : Sentinel primary care influenza-like illness versus severe hospitalized laboratory-confirmed influenza using the moving epidemic method

Monitoring seasonal influenza epidemics is the corner stone to epidemiological surveillance of acute respiratory virus infections worldwide. This work aims to compare two sentinel surveillance systems within the Daily Acute Respiratory Infection Information System of Catalonia (PIDIRAC), the primary care ILI and Influenza confirmed samples from primary care (PIDIRAC-ILI and PIDIRAC-FLU) and the severe hospitalized laboratory confirmed influenza system (SHLCI), in regard to how they behave in the forecasting of epidemic onset and severity allowing for healthcare preparedness. Epidemiological study carried out during seven influenza seasons (2010-2017) in Catalonia, with data from influenza sentinel surveillance of primary care physicians reporting ILI along with laboratory confirmation of influenza from systematic sampling of ILI cases and 12 hospitals that provided data on severe hospitalized cases with laboratory-confirmed influenza (SHLCI-FLU). Epidemic thresholds for ILI and SHLCI-FLU (overall) as well as influenza A (SHLCI-FLUA) and influenza B (SHLCI-FLUB) incidence rates were assessed by the Moving Epidemics Method. Epidemic thresholds for primary care sentinel surveillance influenza-like illness (PIDIRAC-ILI) incidence rates ranged from 83.65 to 503.92 per 100.000 h. Paired incidence rate curves for SHLCI-FLU/PIDIRAC-ILI and SHLCI-FLUA/PIDIRAC-FLUA showed best correlation index' (0.805 and 0.724 respectively). Assessing delay in reaching epidemic level, PIDIRAC-ILI source forecasts an average of 1.6 weeks before the rest of sources paired. Differences are higher when SHLCI cases are paired to PIDIRAC-ILI and PIDIRAC-FLUB although statistical significance was observed only for SHLCI-FLU/PIDIRAC-ILI (p-value Wilcoxon test = 0.039). The combined ILI and confirmed influenza from primary care along with the severe hospitalized laboratory confirmed influenza data from PIDIRAC sentinel surveillance system provides timely and accurate syndromic and virological surveillance of influenza from the community level to hospitalization of severe cases