Targeted antibody and cytokine cancer immunotherapies through collagen affinity

Kazuto Fukunaga, Jeffrey Alan Hubbell, Jun Ishihara, Seung Tae Lee, Lambert Charles François Potin, Michal Mateusz Raczy, Melody Swartz
AMER ASSOC ADVANCEMENT SCIENCE, 2019
Article

Résumé

Cancer immunotherapy with immune checkpoint inhibitors (CPIs) and interleukin-2 (IL-2) has demonstrated clinical efficacy but is frequently accompanied with severe adverse events caused by excessive and systemic immune system activation. Here, we addressed this need by targeting both the CPI antibodies anti-cytotoxic T lymphocyte antigen 4 antibody (alpha CTLA4) + anti-programmed death ligand 1 antibody (alpha PD-L1) and the cytokine IL-2 to tumors via conjugation (for the antibodies) or recombinant fusion (for the cytokine) to a collagen-binding domain (CBD) derived from the blood protein von Willebrand factor (VWF) A3 domain, harnessing the exposure of tumor stroma collagen to blood components due to the leakiness of the tumor vasculature. We show that intravenously administered CBD protein accumulated mainly in tumors. CBD conjugation or fusion decreases the systemic toxicity of both alpha CTLA4+alpha PD-L1 combination therapy and IL-2, for example, eliminating hepatotoxicity with the CPI molecules and ameliorating pulmonary edema with IL-2. Both CBD-CPI and CBD-IL-2 suppressed tumor growth compared to their unmodified forms in multiple murine cancer models, and both CBD-CPI and CBD-IL-2 increased tumor-infiltrating CD8(+) T cells. In an orthotopic breast cancer model, combination treatment with CPI and IL-2 eradicated tumors in 9 of 13 animals with the CBD-modified drugs, whereas it did so in only 1 of 13 animals with the unmodified drugs. Thus, the A3 domain of VWF can be used to improve safety and efficacy of systemically administered tumor drugs with high translational promise.

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https://infoscience.epfl.ch/record/267122?ln=fr

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Kazuto Fukunaga, Jeffrey Alan Hubbell, Jun Ishihara, Seung Tae Lee, Lambert Charles François Potin, Michal Mateusz Raczy, Melody Swartz
AMER ASSOC ADVANCEMENT SCIENCE, 2019
Article

Résumé

Official source

https://infoscience.epfl.ch/record/267122?ln=fr

À propos de ce résultat

Concepts associés (20)

Concepts associés

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Publications associées (36)

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Lung cancer is the leading cause of cancer-associated deaths worldwide. Platinum-based chemotherapy is the most common therapeutic approach in advanced non-small cell lung cancer (NSCLC), particularly for tumors that carry non-druggable activating mutations. However, these tumors are generally not eradicated when diagnosed at an advanced stage and are frequently resistant to chemotherapy. Therefore, there is an urgent medical need for new treatments for this frequent cancer type. Immunotherapies are treatments based on increasing the strength of immune responses against cancer. In particular, monoclonal antibodies that block inhibitory "immune-checkpoints" expressed on T cells (e.g., programmed cell death protein 1, or PD1) show clinical efficacy in increasing tumor types. However, these therapies improve survival in only a minority of NSCLC patients, and ongoing efforts are being made for increasing their efficacy through combinatorial treatments. In this regard, growing evidence indicates that tumor angiogenesis is closely associated with immunosuppression. Pro-angiogenic factors, such as vascular endothelial growth factor A (VEGFA) and angiopoietin 2 (ANG2), cause abnormalities and dysfunctions in tumor blood vessels that impair T cell extravasation and anti-tumoral activity. Recent findings in preclinical cancer models show that reprogramming the features of tumor-associated blood vessels by anti-angiogenic drugs may help to increase intratumoral T cell abundance and sensitize refractory tumors to immunotherapy. Nevertheless, the potential of anti-angiogenics in the context of lung cancer immunotherapy is poorly understood. Therefore, the main objective of my project was to audit combinations of anti-angiogenic therapy and immune checkpoint blockade in a genetically engineered mouse model of NSCLC. I first studied the anti-tumoral activity of dual VEGFA and ANG2 inhibition using a bispecific antibody (A2V) in the KrasLSL-G12D/+;p53fl/fl (KP) NSCLC model. I found that A2V, but not single VEGFA or ANG2 blockade, had robust tumor-inhibitory activity in KP mice, which was at least comparable to standard-of-care chemotherapy. However, A2V only moderately increased cytotoxic T cells in the tumors. To potentially increase the immunogenicity of KP tumors, I also generated two KP mouse variants by either co-expressing a surrogate neoantigen or genetically deleting a DNA mismatch repair enzyme in the cancer cells. However, the therapeutic efficacy of A2V was not further improved in spite of a slightly more activated immune response. I next investigated combination therapies involving A2V and PD1 blockade. In contrast to previous findings in other cancer types, the addition of PD1 antibodies deteriorated the efficacy of A2V, likely through enhancing immunosuppressive mechanisms sustained, at least in part, by regulatory T cells and T cell-antagonizing cytokines. In summary, my work supports the potential clinical efficacy of combined VEGFA and ANG2 blockade for lung cancer therapy but provided little evidence for its synergy in association with anti-PD1 therapy.

EPFL2019

Chargement

The identification of cancer stem cells (CSCs) has fundamentally changed the understanding of tumor biology. CSCs drive tumor initiation and long term tumor growth, can cause relapse and are essential for metastatic colonization. These features place cancer stem cells in the focus for cancer therapy. Recently, great advances have been made in the field of cancer adoptive immunotherapy using engineered T cells. Combining both fields of cancer research to generate immune therapies directly targeting CSCs for increased efficacy and long-term anti-tumor effects would generate great hope for cancer patients. Herein, proof of concept is made that chimeric antigen receptor (CARs) T cells killing breast CSCs are capable of eliminating established metastases. Killing of the CSCs modifies the immune environment resulting in the loss of regulatory T cells and in the induction of an endogenous immune response directed against the tumor that is indispensable for treatment success. Antibody mediated ablation of the CAR T cells induced a rapid re-appearance of CSCs and re-establishment of an immunosuppressive environment in the tumor creating a need for continuous targeting of the CSCs. I demonstrate that endogenous T cell responses are less effective against CSCs than the rest of the tumor. Furthermore, proof is made across multiple tumor models, that activated immune cells and inflammation induce dedifferentiation of bulk tumor cells which regain stem-like phenotypes. This observation further emphasizes the importance of directly targeting the CSCs. Studying the interplay between CSCs and the immune system requires syngeneic, immune competent murine models. This greatly limits the transgenic markers that one can use to visualize, track and target CSCs. Herein, we developed different tolerant mouse models to study breast cancer stem cells in vivo. 4T1 breast adenocarcinoma cells were engineered to express hCEA or GFP-based model surface antigens under the control a stem cell specific promoter. These cells, injected into syngeneic model antigen-tolerant mice established lung metastases which were eliminated by CAR T cell therapy. To further validate this approach using endogenous CSC markers, a genetic cancer model (MMTV-PyMT) has been developed, where stem cell Identification is based on CD90 allelic differentiation. Overall, this work highlights an important interplay between CSCs and the immune system. My results suggest that a feedback loop may be created were immune attack induces CSCs, which are immune privileged and escape killing from endogenous T cells. Subsequently, CSCs induce and control an immunosuppressive environment leading to immune escape. This provides a strong rationale to develop CSC-targeted therapies to be administered in combination with immunotherapies.

EPFL2017

Chargement

An autonomous microfluidic device for identification and analysis of individual clinically-relevant mammalian cells

Fabien Louis Claude Robert Jammes

Cancer represents one of the major public health challenges worldwide, representing more than 25% of all deaths in the European Union in 2014. It affects a large and growing part of the general population, with the National Cancer Institute (NCI) estimating that nearly 40% of men and women will be diagnosed with cancer at some point in their lifetimes.Cancer presents an enormous intrinsic complexity and diversity as well as a pronounced resistance to interventions. Current treatments are composed primarily of chemotherapy, surgery, radiotherapy and drug therapy. Each of these approaches has been met with restrained successes and limitations. One growing field of cancer treatments that is showing promising results is immunotherapy. An improved understanding of the cancer biology and microenvironment over the years, particularly the interactions between a tumor and the immune system, has demonstrated the capability of the host immune system to detect and eliminate malignant cells through the process of immune surveillance. However, by various mechanisms, malignant cells can stochastically "hide" from the immune system and transform into tumors. Immunotherapies intend to use the immune system to fight the tumor progression. Different kinds of immunotherapies exist: from immune checkpoint inhibitors (PD-1), to monoclonal antibody treatments and cancer vaccines. One interesting approach of immunotherapy is called adoptive cell transfer therapy (ACT), wherein immune cells, and more precisely, T cells, from the cancer-affected patient are extracted, allowing tumor-specific and effective T cells to be isolated, expanded, activated and re-injected into the patient's body. This type of treatment has already demonstrated great tumor clearance potential as well as the ability to induce long-term cancer immunity. However, one of the major challenges in this type of treatment is the recognition, isolation and expansion of high-affinity, effective T cells that are able to eliminate cancer cells. Several key advances in the field of genetics, sequencing, and personalized medicine have allowed for the identification of tumor-specific antigens, the unique cancer signature for a given patient and a specific tumor. But there is a growing need for novel tools and platforms enabling the screening, selection and isolation of rare, naturally-occurring, high-affinity, tumor-specific T cells. In this thesis we present a novel approach to study the affinity of CD8+ lymphocytes at the single-cell level using microfluidics technology. We demonstrate the ability of our platform to isolate single cells and evaluate their affinities towards a given antigen by using the reversible pMHC NTAmers technology developed by Julien Schmidt et al. at the Ludwig Institute for Cancer Research in Lausanne, Switzerland. The innovative platform developed here effectively alleviates some of the bottlenecks of current technologies, like flow cytometry, for affinity studies of lymphocytes at the single-cell level. This thesis represents the development from idea to proof-of-concept of this novel platform and we herein describe the origins and characterization of this technology as well as the first applications towards a personalized-medicine tool for clinical research uses.

EPFL2021

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EPFL2019

EPFL2017

An autonomous microfluidic device for identification and analysis of individual clinically-relevant mammalian cells

Fabien Louis Claude Robert Jammes

EPFL2021