Immunomodulatory mAbs as Tools to Investigate on Cis-Interaction of PD-1/PD-L1 on Tumor Cells and to Set Up Methods for Early Screening of Safe and Potent Combinatorial Treatments

Antibodies targeting Immune Checkpoints (IC) on tumor infiltrating lymphocytes improve immune responses against cancer. Recently, the expression of some ICs has also been reported on cancer cells. We used the clinically validated Ipilimumab and Nivolumab and other novel human antibodies targeting Cytotoxic T- lymphocyte-antigen 4 (CTLA-4), Programmed Death receptor-1 (PD-1) and Programmed Death Ligand 1 (PD-L1) to shed light on the functions of these ICs in cancer cells. We show here for the first time that all these antagonistic mAbs are able to reduce Erk phosphorylation and, unexpectedly, to induce a significant increase of ICs expression on tumor cells, involving a hyperphosphorylation of NF-kB. On the contrary, agonistic PD-L1 and PD-1 recombinant proteins showed opposite effects by leading to a significant reduction of PD-1 and PD-L1, thus also suggesting the existence of a crosstalk in tumor cells between multiple ICs. Since the immunomodulatory mAbs show their higher anti-tumor efficacy by activating lymphocytes against cancer cells, we also investigated whether it was possible to identify the most efficient combinations of immunomodulatory mAbs for achieving potent anti-tumor efficacy associated with the lowest adverse side effects by setting up novel simple and predictive in vitro models based on co-cultures of tumor cells or human fetal cardiomyocytes with lymphocytes. We demonstrate here that novel combinations of immunomodulatory mAbs with more potent anti-cancer activity than Ipilimumab and Nivolumab combination can be identified with no or lower cardiotoxic side effects. Thus, we propose these co-cultures-based assays as useful tools to test also other combinatorial treatments of emerging immunomodulatory mAbs against different ICs for the early screening of most potent and safe combinatorial therapeutic regimens.

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Recent Advances and Future Prospects in Immune Checkpoint (ICI)-Based Combination Therapy for Advanced HCC

Cancers ◽

10.3390/cancers13081949 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1949

Author(s):

Yawen Dong ◽

Jeffrey Sum Lung Wong ◽

Ryohichi Sugimura ◽

Ka-On Lam ◽

Bryan Li ◽

...

Keyword(s):

Immune Checkpoint ◽

Cytotoxic T Lymphocyte ◽

Checkpoint Inhibitors ◽

Immune Checkpoints ◽

Advanced Hcc ◽

Oncogenic Pathways ◽

Vegf Pathway ◽

And Function ◽

Endothelial Growth Factor

Advanced, unresectable hepatocellular carcinoma has a dismal outcome. Multiple immune checkpoint inhibitors (ICIs) targeting the programmed-cell death 1 pathway (PD-1/L1) have been approved for the treatment of advanced HCC. However, outcomes remain undesirable and unpredictable on a patient-to-patient basis. The combination of anti-PD-1/L1 with alternative agents, chiefly cytotoxic T-lymphocyte antigen-4 (CTLA-4) ICIs or agents targeting other oncogenic pathways such as the vascular endothelial growth factor (VEGF) pathway and the c-MET pathway, has, in addition to the benefit of directly targeting alterative oncogenic pathways, in vitro evidence of synergism through altering the genomic and function signatures of T cells and expression of immune checkpoints. Several trials have been completed or are underway evaluating such combinations. Finally, studies utilizing transcriptomics and organoids are underway to establish biomarkers to predict ICI response. This review aims to discuss the biological rationale and clinical advances in ICI-based combinations in HCCs, as well as the progress and prospects of the search for the aforementioned biomarkers in ICI treatment of HCC.

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Patterns and Relevance of Langerhans Islet Invasion in Pancreatic Cancer

Cancers ◽

10.3390/cancers13020249 ◽

2021 ◽

Vol 13 (2) ◽

pp. 249

Author(s):

Ruediger Goess ◽

Ayse Ceren Mutgan ◽

Umut Çalışan ◽

Yusuf Ceyhun Erdoğan ◽

Lei Ren ◽

...

Keyword(s):

Diabetes Mellitus ◽

Pancreatic Cancer ◽

Cancer Cells ◽

Tumor Cells ◽

Islet Cells ◽

Human Pancreatic Cancer ◽

Type I ◽

Type Iv ◽

Pancreatic Cancer Cells

Background: Pancreatic cancer‐associated diabetes mellitus (PC‐DM) is present in most patients with pancreatic cancer, but its pathogenesis remains poorly understood. Therefore, we aimed to characterize tumor infiltration in Langerhans islets in pancreatic cancer and determine its clinical relevance. Methods: Langerhans islet invasion was systematically analyzed in 68 patientswith pancreatic ductal adenocarcinoma (PDAC) using histopathological examination and 3D in vitro migration assays were performed to assess chemoattraction of pancreatic cancer cells to isletcells. Results: Langerhans islet invasion was present in all patients. We found four different patterns of islet invasion: (Type I) peri‐insular invasion with tumor cells directly touching the boundary, but not penetrating the islet; (Type II) endo‐insular invasion with tumor cells inside the round islet; (Type III) distorted islet structure with complete loss of the round islet morphology; and (Type IV)adjacent cancer and islet cells with solitary islet cells encountered adjacent to cancer cells. Pancreatic cancer cells did not exhibit any chemoattraction to islet cells in 3D assays in vitro. Further, there was no clinical correlation of islet invasion using the novel Islet Invasion Severity Score (IISS), which includes all invasion patterns with the occurrence of diabetes mellitus. However, Type IV islet invasion was related to worsened overall survival in our cohort. Conclusions: We systematically analyzed, for the first time, islet invasion in human pancreatic cancer. Four different main patterns of islet invasion were identified. Diabetes mellitus was not related to islet invasion. However, moreresearch on this prevailing feature of pancreatic cancer is needed to better understand underlying principles.

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Nanostructured Dihydroartemisinin Plus Epirubicin Liposomes Enhance Treatment Efficacy of Breast Cancer by Inducing Autophagy and Apoptosis

Nanomaterials ◽

10.3390/nano8100804 ◽

2018 ◽

Vol 8 (10) ◽

pp. 804 ◽

Cited By ~ 3

Author(s):

Ying-Jie Hu ◽

Jing-Ying Zhang ◽

Qian Luo ◽

Jia-Rui Xu ◽

Yan Yan ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Therapeutic Strategy ◽

Beclin 1 ◽

Type I ◽

Programmed Death ◽

Cancer Tissues

The heterogeneity of breast cancer and the development of drug resistance are the relapse reasons of disease after chemotherapy. To address this issue, a combined therapeutic strategy was developed by building the nanostructured dihydroartemisinin plus epirubicin liposomes. Investigations were performed on human breast cancer cells in vitro and xenografts in nude mice. The results indicated that dihydroartemisinin could significantly enhance the efficacy of epirubicin in killing different breast cancer cells in vitro and in vivo. We found that the combined use of dihydroartemisinin with epirubicin could efficiently inhibit the activity of Bcl-2, facilitate release of Beclin 1, and further activate Bax. Besides, Bax activated apoptosis which led to the type I programmed death of breast cancer cells while Beclin 1 initiated the excessive autophagy that resulted in the type II programmed death of breast cancer cells. In addition, the nanostructured dihydroartemisinin plus epirubicin liposomes prolonged circulation of drugs, and were beneficial for simultaneously delivering drugs into breast cancer tissues. Hence, the nanostructured dihydroartemisinin plus epirubicin liposomes could provide a new therapeutic strategy for treatment of breast cancer.

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Synergy of nanodiamond-doxorubicin conjugates and PD-L1 blockade effectively turns tumor associated macrophages against tumor cells

10.21203/rs.3.rs-715564/v1 ◽

2021 ◽

Author(s):

Huazhen Xu ◽

Tongfei Li ◽

Chao Wang ◽

Yan Ma ◽

Yan Liu ◽

...

Keyword(s):

Lung Cancer ◽

Tumor Cell ◽

Cancer Cells ◽

Tumor Cells ◽

Lung Cancer Cells ◽

Dependent Manner ◽

Tumor Associated Macrophages ◽

M1 Type

Abstract Background: Tumor-associated macrophages (TAM) are the most abundant stromal cells in the tumor microenvironment. Turning the TAM against their host tumor cells is an intriguing therapeutic strategy particularly attractive for patients with immunologically “cold” tumors. This concept was mechanistically demonstrated on in vitro human and murine lung cancer cells and their corresponding TAM models through combinatorial use of nanodiamond-doxorubicin conjugates (Nano-DOX) and a PD-L1 blocking agent BMS-1. Nano-DOX are an agent previously proved to be able to stimulate tumor cells’ immunogenicity and thereby reactivate the TAM into the anti-tumor M1 phenotype. Results: Nano-DOX were first shown to stimulate the tumor cells and the TAM to release the cytokine HMGB1 which, regardless of its source, acted through the RAGE/NF-κB pathway to induce PD-L1 in the tumor cells and PD-L1/PD-1 in the TAM. Interestingly, Nano-DOX also induced NF-κB-dependent RAGE expression in the tumor cells and thus reinforced HMGB1’s action thereon. Then, BMS-1 was shown to enhance Nano-DOX-stimulated M1-type activation of TAM both by blocking Nano-DOX-induced PD-L1 in the TAM and by blocking tumor cell PD-L1 ligation with TAM PD-1. The TAM with enhanced M1-type repolarization both killed the tumor cells and suppressed their growth. BMS-1 could also potentiate Nano-DOX’s action to suppress tumor cell growth via blocking of Nano-DOX-induced PD-L1 therein. Finally, Nano-DOX and BMS-1 achieved synergistic therapeutic efficacy against in vivo tumor grafts in a TAM-dependent manner. Conclusions: PD-L1/PD-1 upregulation mediated by autocrine and paracrine activation of the HMGB1/RAGE/NF-κB signaling is a key response of lung cancer cells and their TAM to stress, which can be induced by Nano-DOX. Blockade of Nano-DOX-induced PD-L1, both in the cancer cells and the TAM, achieves enhanced activation of TAM-mediated anti-tumor response.

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Fusion Hybrid of Dendritic Cells and Engineered Tumor Cells Expressing Interleukin-12 Induces Type 1 Immune Responses against Tumor

Tumori Journal ◽

10.1177/030089160509100614 ◽

2005 ◽

Vol 91 (6) ◽

pp. 531-538 ◽

Cited By ~ 11

Author(s):

Meiqing Shi ◽

Liping Su ◽

Sigou Hao ◽

Xulin Guo ◽

Jim Xiang

Keyword(s):

Cancer Immunotherapy ◽

Tumor Cells ◽

Antitumor Immunity ◽

Cytotoxic T Lymphocyte ◽

Tumor Regression ◽

Interleukin 12 ◽

Myeloma Cells ◽

Th1 Cytokine

Aims and Background Dendritic cell (DC)-tumor fusion hybrid vaccinees that facilitate antigen presentation represent a novel powerful strategy in cancer immunotherapy. Preclinical studies have demonstrated that IL-12 promotes specific antitumor immunity mediated by T cells in several types of tumors. In the present study, we investigated the antitumor immunity derived from vaccination of fusion hybrids between DCs and engineered J558/IL-12 myeloma cells secreting Th1 cytokine IL-12. Methods The expression vector pcDNA-IL-12 was generated and transfected into J558 myeloma cells and then bone marrow-derived DCs were fused with engineered J558/IL-12 cells. The antitumor immunity derived from vaccination of the fusion hybrid DC/J558/IL-12 was evaluated in vitro and in vivo. Results DC/J558/IL-12 cells secreted recombinant IL-12 (1.6 ng/mL), and inoculation of BALB/c mice with DC/J558/IL-12 hybrid induced a Th1 dominant immune response and resulted in tumor regression. Immunization of mice with engineered DC/J558/IL-12 hybrid elicited stronger J558 tumor-specific cytotoxic T lymphocyte (CTL) responses in vitro as well as more potent protective immunity against J558 tumor challenge in vivo than immunization with the mixture of DCs and J558/IL-12, J558/IL-12 and J558, respectively. Furthermore, the antitumor immunity mediated by DC/J558/1L-12 tumor cell vaccination in vivo appeared to be dependent on CD8+ CTL. Conclusions These results demonstrate that the engineered fusion hybrid vaccines that combine Th1 cytokine gene-modified tumor cells with DCs may be an attractive strategy for cancer immunotherapy.

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A Nanobody Against Cytotoxic T-Lymphocyte Associated Antigen-4 Increases the Anti-Tumor Effects of Specific CD8+ T Cells

Journal of Biomedical Nanotechnology ◽

10.1166/jbn.2019.2859 ◽

2019 ◽

Vol 15 (11) ◽

pp. 2229-2239 ◽

Cited By ~ 3

Author(s):

Zhuoran Tang ◽

Fengzhen Mo ◽

Aiqun Liu ◽

Siliang Duan ◽

Xiaomei Yang ◽

...

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Cell ◽

Tumor Cells ◽

Cytotoxic T Lymphocyte ◽

Ex Vivo ◽

T Cell Proliferation ◽

Tumor Cell Apoptosis ◽

Xenograft Models

Adoptive cell-based immunotherapy typically utilizes cytotoxic T lymphocytes (CTLs), expanding these cells ex vivo. Such expansion is traditionally accomplished through the use of autologous APCs that are capable of interactions with T cells. However, incidental inhibitory program such as CTLA-4 pathway can impair T cell proliferation. We therefore designed a nanobody which is specific for CTLA-4 (CTLA-4 Nb 16), and we then used this molecule to assess its ability to disrupt CTLA-4 signaling and thereby overcome negative costimulation of T cells. With CTLA-4 Nb16 stimulation, dendritic cell/hepatocellular carcinoma fusion cells (DC/HepG2-FCs) enhanced autologous CD8+ T cell proliferation and production of IFN-γ in vitro, thereby leading to enhanced killing of tumor cells. Using this approach in the context of adoptive CD8+ immunotherapy led to a marked suppression of tumor growth in murine NOD/SCID hepatocarcinoma or breast cancer xenograft models. We also observed significantly increased tumor cell apoptosis, and corresponding increases in murine survival. These findings thus demonstrate that in response to nanobody stimulation, DC/tumor cells-FC-induced specific CTLs exhibit superior anti-tumor efficacy, making this a potentially valuable means of achieving better adoptive immunotherapy outcomes in cancer patients.

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Low dose amiodarone reduces tumor growth and angiogenesis

Scientific Reports ◽

10.1038/s41598-020-75142-1 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Eliana Steinberg ◽

Arnon Fluksman ◽

Chalom Zemmour ◽

Katerina Tischenko ◽

Adi Karsch-Bluman ◽

...

Keyword(s):

Side Effects ◽

Cancer Cells ◽

Low Dose ◽

Tumor Model ◽

In Vitro Assays ◽

Potential Candidate ◽

Key Factor ◽

Angiogenic Effect

Abstract Amiodarone is an anti-arrhythmic drug that was approved by the US Food and Drug Administration (FDA) in 1985. Pre-clinical studies suggest that Amiodarone induces cytotoxicity in several types of cancer cells, thus making it a potential candidate for use as an anti-cancer treatment. However, it is also known to cause a variety of severe side effects. We hypothesized that in addition to the cytotoxic effects observed in cancer cells Amiodarone also has an indirect effect on angiogensis, a key factor in the tumor microenvironment. In this study, we examined Amiodarone's effects on a murine tumor model comprised of U-87 MG glioblastoma multiforme (GBM) cells, known to form highly vascularized tumors. We performed several in vitro assays using tumor and endothelial cells, along with in vivo assays utilizing three murine models. Low dose Amiodarone markedly reduced the size of GBM xenograft tumors and displayed a strong anti-angiogenic effect, suggesting dual cancer fighting properties. Our findings lay the ground for further research of Amiodarone as a possible clinical agent that, used in safe doses, maintains its dual properties while averting the drug’s harmful side effects.

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Generation of cytotoxic immune responses during the progression of a rat glioma

Journal of Neurosurgery ◽

10.3171/jns.1994.80.1.0090 ◽

1994 ◽

Vol 80 (1) ◽

pp. 90-96 ◽

Cited By ~ 19

Author(s):

Frank P. Holladay ◽

Rajani Choudhuri ◽

Teresa Heitz ◽

Gary W. Wood

Keyword(s):

Immune Responses ◽

Tumor Progression ◽

Tumor Cells ◽

T Lymphocyte ◽

Cytotoxic T Lymphocyte ◽

Interleukin 2 ◽

Autologous Tumor ◽

Content Type ◽

Tumor Associated Antigens

✓ Cytotoxic T lymphocytes specific for tumor-associated antigens are produced by exposing animals to tumor cells and stimulating lymphocytes from animals immunized in vitro with tumor cells and small amounts of interleukin-2 (IL-2). This study was designed to determine whether a fast-growing immunogenic avian sarcoma virus-induced glioma produces primed cytotoxic T lymphocyte precursors during its progression. Lymphocytes from intracerebral glioma-bearing rats generally failed to proliferate in vitro in response to immunization with tumor cells and IL-2 and, when proliferative responses were observed, the lymphocytes were not cytotoxic for glioma cells. However, when the same tumor was growing subcutaneously, lymphocytes proliferated and exhibited glioma-specific cytotoxicity when stimulated in vitro with autologous tumor cells and IL-2. Subcutaneous immunization of intracerebral glioma-bearing rats with tumor cells and adjuvant induced strong cytotoxic T lymphocyte responses. The results demonstrated that, while intracerebral tumor progression itself does not induce an antiglioma immune response, immune responses to tumor-associated antigens may be induced by systemic immunization of tumor-bearing animals. The results suggest that the immunogenicity of brain tumors is masked by the immunologically privileged status of the brain, not by the induction of generalized immune suppression during tumor progression.

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Improved in vivo efficacy of tumor-infiltrating lymphocytes after restimulation with irradiated tumor cells in vitro

Annals of Surgical Oncology ◽

10.1007/bf02306093 ◽

1996 ◽

Vol 3 (6) ◽

pp. 580-587 ◽

Cited By ~ 8

Author(s):

Ulrike L. Burger ◽

Maximilian P. Chang ◽

Makoto Nagoshi ◽

Peter S. Goedegebuure ◽

Timothy J. Eberlein

Keyword(s):

Tumor Cells ◽

Tumor Infiltrating Lymphocytes ◽

In Vivo Efficacy ◽

Infiltrating Lymphocytes

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Development of Self-Assembled Biomimetic Boc-Protected Peptide-Polymer Based Nanovehicles for Targeted Delivery to Tumor Cells

Journal of Biomimetics Biomaterials and Biomedical Engineering ◽

10.4028/www.scientific.net/jbbbe.29.33 ◽

2016 ◽

Vol 29 ◽

pp. 33-53 ◽

Cited By ~ 1

Author(s):

Alexandra M. Brown ◽

Yoliem S. Miranda-Alarćon ◽

Grant A. Knoll ◽

Steven M. Romanelli ◽

Ipsita A. Banerjee

Keyword(s):

Breast Cancer ◽

Side Effects ◽

Cancer Cells ◽

Tumor Cells ◽

Targeted Delivery ◽

Breast Cancer Cells ◽

Dermal Fibroblasts ◽

Burst Release ◽

Chemotherapeutic Drugs ◽

New Family

Although effective, chemotherapeutic drugs often cause undesired side-effects. Thus, encapsulating chemotherapeutic drugs into nanoscale drug delivery vehicles (DDVs) has the potential to reduce side effects and promote targeted delivery. By mimicking ABA like block-co-polymer systems, we have developed a new amphiphilic biomimetic co-polymer Boc-Ile-PEG-Ile-Boc which was found to readily self-assemble into nanomicelles within hydrophilic shell structures. To facilitate targeting tumor cells, the nanoassemblies were bound to folate, leading to the formation of core shell like structures (IBP-F). Gold nanoparticles (AuNPs), were then embedded followed by functionalization with a second layer of folate. The final DDV system abbreviated (IBP-F-Au-F) formed a multi-layered nanostructure that was capable of efficiently encapsulating the anti-tumor drug tamoxifen. For comparison, we also examined the efficacy of the IBP-F assemblies as DDVs in the absence of AuNPs and a second folate layer. Release profiles showed an initial burst release, followed by sustained release. The DDVs were found to be biocompatible. Upon encapsulating the DDVs with tamoxifen, cell proliferation was inhibited over a period of 72 hours for both DDVs, while non-cancerous dermal fibroblasts continued to proliferate, thus indicating specific targeting ability of the DDVs. Confocal microscopy studies conducted in the presence of human breast cancer cells, MDA-MDB 231 revealed that the drug loaded assemblies were successfully internalized within the cells. SPR analysis demonstrate that IBP-F-Au-F had a higher affinity for breast cancer cells over non-cancerous keratinocyte cells. Thus, we have developed a new family of DDVs that selectively targets tumor cells.

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