The use of monoclonal antibodies (mAbs) for cancer therapy has achieved considerable success in recent years. Antibody-drug conjugates are powerful new treatment options for lymphomas and solid tumours, and immunomodulatory antibodies have also recently achieved remarkable clinical success. The development of therapeutic antibodies requires a deep understanding of cancer serology, protein-engineering techniques, mechanisms of action and resistance, and the interplay between the immune system and cancer cells. This Review outlines the fundamental strategies that are required to develop antibody therapies for cancer patients through iterative approaches to target and antibody selection, extending from preclinical studies to human trials.

https://www.nature.com/articles/nrc3236.pdf?proof=t

Antibody therapy of cancer

With more than 20 molecules in clinical use, monoclonal antibodies have finally come of age as therapeutics, generating a market value of $11 billion in 2004, expected to reach $26 billion by 2010. While delivering interesting results in the treatment of several major diseases including autoimmune, cardiovascular and infectious diseases, cancer and inflammation, clinical trials and research are generating a wealth of useful information, for instance about associations of clinical responses with Fc receptor polymorphisms and the infiltration and recruitment of effector cells into targeted tissues. Some functional limitations of therapeutic antibodies have come to light such as inadequate pharmacokinetics and tissue accessibility as well as impaired interactions with the immune system, and these deficiencies point to areas where additional research is needed. This review aims at giving an overview of the current state of the art and describes the most promising avenues that are being followed to create the next generation of antibody-based therapeutic agents.

This article is part of a themed section on Vector Design and Drug Delivery. For a list of all articles in this section see the end of this paper, or visit: http://www3.interscience.wiley.com/journal/121548564/issueyear?year=2009

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Therapeutic antibodies: successes, limitations and hopes for the future.

The human epidermal growth factor receptor (HER-2) oncogene encodes a transmembrane tyrosine kinase receptor that has evolved as a major classifier of invasive breast cancer and target of therapy for the disease. The validation of the general prognostic significance of HER-2 gene amplification and protein overexpression in the absence of anti-HER-2 targeted therapy is discussed in a study of 107 published studies involving 39,730 patients, which produced an overall HER-2-positive rate of 22.2% and a mean relative risk for overall survival (OS) of 2.74. The issue of HER-2 status in primary versus metastatic breast cancer is considered along with a section on the features of metastatic HER-2-positive disease. The major marketed slide-based HER-2 testing approaches, immunohistochemistry, fluorescence in situ hybridization, and chromogenic in situ hybridization, are presented and contrasted in detail against the background of the published American Society of Clinical Oncology-College of American Pathologists guidelines for HER-2 testing. Testing issues, such as the impact of chromosome 17 polysomy and local versus central HER-2 testing, are also discussed. Emerging novel HER-2 testing techniques, including mRNA-based testing by real-time polymerase chain reaction and DNA microarray methods, HER-2 receptor dimerization, phosphorylated HER-2 receptors, and HER-2 status in circulating tumor cells, are also considered. A series of biomarkers potentially associated with resistance to trastuzumab is discussed with emphasis on the phosphatase and tensin homologue deleted on chromosome ten/Akt and insulin-like growth factor receptor pathways. The efficacy results for the more recently approved small molecule HER-1/HER-2 kinase inhibitor lapatinib are also presented along with a more limited review of markers of resistance for this agent. Additional topics in this section include combinations of both anti-HER-2 targeted therapies together as well as with novel agents including bevacizumab, everolimus, and tenespimycin. A series of novel HER-2-targeting agents is also presented, including pertuzumab, ertumaxomab, HER-2 vaccines, and recently discovered tyrosine kinase inhibitors. Biomarkers predictive of HER-2 targeted therapy toxicity are included, and the review concludes with a consideration of HER-2 status in the prediction of response to non-HER-2 targeted treatments including hormonal therapy, anthracyclines, and taxanes.

The HER-2 Receptor and Breast Cancer: Ten Years of Targeted Anti–HER-2 Therapy and Personalized Medicine

Aberrant receptor expression or functioning of the epidermal growth factor receptor (Erbb) family plays a crucial part in the development and evolution of cancer. Inhibiting the signalling activity of individual receptors in this family has advanced the treatment of a range of human cancers. In this Review we re-evaluate the role of two important family members, ERBB2 (also known as HER2) and ERBB3 (also known as HER3), and explore the mechanisms of action and preclinical and clinical data for new therapies that target signalling through these pivotal receptors. These new therapies include tyrosine kinase inhibitors, antibody-chemotherapy conjugates, heat-shock protein inhibitors and antibodies that interfere with the formation of ERBB2-ERBB3 dimers.

http://dipbsf.uninsubria.it/monti/BFPN%202010/Biblio%20BFPN%202010/BFPN%20RTKs/ERBB2%20&%20ERBB3.pdf

Novel anticancer targets: revisiting ERBB2 and discovering ERBB3

It has been more than three decades since the first monoclonal antibody was approved by the United States Food and Drug Administration (US FDA) in 1986, and during this time, antibody engineering has dramatically evolved. Current antibody drugs have increasingly fewer adverse effects due to their high specificity. As a result, therapeutic antibodies have become the predominant class of new drugs developed in recent years. Over the past five years, antibodies have become the best-selling drugs in the pharmaceutical market, and in 2018, eight of the top ten bestselling drugs worldwide were biologics. The global therapeutic monoclonal antibody market was valued at approximately US$115.2 billion in 2018 and is expected to generate revenue of $150 billion by the end of 2019 and $300 billion by 2025. Thus, the market for therapeutic antibody drugs has experienced explosive growth as new drugs have been approved for treating various human diseases, including many cancers, autoimmune, metabolic and infectious diseases. As of December 2019, 79 therapeutic mAbs have been approved by the US FDA, but there is still significant growth potential. This review summarizes the latest market trends and outlines the preeminent antibody engineering technologies used in the development of therapeutic antibody drugs, such as humanization of monoclonal antibodies, phage display, the human antibody mouse, single B cell antibody technology, and affinity maturation. Finally, future applications and perspectives are also discussed.

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Development of therapeutic antibodies for the treatment of diseases

PROBLEM TO BE SOLVED: To obtain human or animal tumor cells that express an antibody to the surface antigen on the antigen-presenting cells(APC), have no immunoglobulin originating from the parent tumor, can be applied to not only B-cell tumors but also to other tumors and is useful for producing antitumor agent. SOLUTION: The objective tumor cells express the antibody to the surface antigen on the APC and have no immunoglobulin originating from the parent tumor. The anti-APC antibody bonds to the membrane and the tumor cells for example, epithelial tumor cells, mixed mesenchymal tumor, hematopoietic tumor cells and the like are obtained by introducing the immunoglobulin encoding the anti-APC antibody or by fusing with the hybridoma cells producing the anti-APC antibody. In a preferred embodiment, the tumor cells originating from the patient are dosed to the tumor patient to allow the tumor cells expressing the anti-APC antibody to the APC to turn to the APC again thereby inducing the anti-tumor immunization.

Induction of antitumor immunity by inducing tumor cells to antigen-presenting cells again

Procédé d'immunisation ex vivo utilisant des anticorps hétérologues intactes bispécifiques ou trispécifiques

Introduction: Peritoneal carcinomatosis of solid tumor is a fatal tumor diagnosis without standard treatment. In a pilot study, a new therapy concept using trifunctional antibodies (trAb) was evaluated, which are able to redirect CD3+ T-lymphocytes to tumor cells and to induce anti-tumor immunity by activation of Fcγ-receptor type I and III accessory cells by their intact Fc fragment. Material and methods: In 9 patients (group A) with peritoneal carcinomatosis of solid tumors, intraperitoneal application of trAb was performed after tumor resection and/or ineffective chemotherapy. Additionally, 8 patients with malignant ascites due to peritoneal carcinomatosis (group B) were treated intraperitoneally. Treatment consisted of 2–6 applications of the trAbs anti-EpCAM × anti-CD3 or anti-HER2/neu × anti-CD3 (30–940) µg within 9–23 days. In group A, restimulation by trAb + PBMC + irradiated autologous tumor cells was performed after 30 days. After another 10 days, specific tumor reactive T-lymphocytes were detected by FACS analysis. In group B, monitoring of tumor cells in ascites was done by immunofluorescence staining, FACS and PCR analysis. Results: i. p. treatment was tolerated without severe side effects. In group A, 5/9 patients had tumor reactive T-lymphocytes after therapy and restimulation. 5/9 patients showed a clinical response (stable disease, partial remission) with time to progression of 3.6 months. In group B, ascites accumulation was diminished in 8/8 patients, which was correlated to a total elimination of tumor cells in ascites (p = 0.04). Conclusion: Intraperitoneal application of trifunctional antibodies showed convincing immunological effects, as induction of active anti-tumor immunity and clinical efficacy was seen in patients with peritoneal carcinomatosis. The novel concept is actually evaluated in clinical phase II/III trials.

Intraperitoneale Applikation trifunktioneller Antikörper: Ein neuartiges Konzept zur Behandlung der Peritonealkarzinose solider Tumoren

Introduction Despite advances in treating high-risk neuroblastoma, 50-60% of patients still suffer relapse, necessitating new treatment options. Bispecific trifunctional antibodies (trAbs) are a promising new class of immunotherapy. TrAbs are heterodimeric IgG-like molecules that bind CD3 and a tumor-associated antigen simultaneously, whereby inducing a TCR-independent anti-cancer T cell response. Moreover, via their functional Fc region they recruit and activate cells of the innate immune system like antigen-presenting cells potentially enhancing induction of adaptive tumor-specific immune responses. Methods We used the SUREK trAb, which is bispecific for GD2 and murine Cd3. Tumor-blind trAb and the monoclonal ch14.18 antibody were used as controls. A co-culture model of murine dendritic cells (DCs), T cells and a neuroblastoma cell line was established to evaluate the cytotoxic effect and the T cell effector function in vitro. Expression of immune checkpoint molecules on tumor-infiltrating T cells and the induction of an anti-neuroblastoma immune response using a combination of whole cell vaccination and trAb therapy was investigated in a syngeneic immunocompetent neuroblastoma mouse model (NXS2 in A/J background). Finally, vaccinated mice were assessed for the presence of neuroblastoma-directed antibodies. We show that SUREK trAb-mediated effective killing of NXS2 cells in vitro was strictly dependent on the combined presence of DCs and T cells. Results Using a syngeneic neuroblastoma mouse model, we showed that vaccination with irradiated tumor cells combined with SUREK trAb treatment significantly prolonged survival of tumor challenged mice and partially prevent tumor outgrowth compared to tumor vaccination alone. Treatment led to upregulation of programmed cell death protein 1 (Pd-1) on tumor infiltrating T cells and combination with anti-Pd-1 checkpoint inhibition enhanced the NXS2-directed humoral immune response. Conclusion Here, we provide first preclinical evidence that a tumor vaccination combined with SUREK trAb therapy induces an endogenous anti-neuroblastoma immune response reducing tumor recurrence. Furthermore, a combination with anti-Pd-1 immune checkpoint blockade might even further improve this promising immunotherapeutic concept in order to prevent relapse in high-risk neuroblastoma patients.

Combination of GD2-directed bispecific trifunctional antibody therapy with Pd-1 immune checkpoint blockade induces anti-neuroblastoma immunity in a syngeneic mouse model

Die Erfindung beschreibt die Verwendung trifunktioneller bispezifischer und trispezifischer Antikorper zur Zerstorung von Tumorzellen, die z. B. durch einen chirurgischen Eingriff intraperitoneal disseminiert werden, wobei der Antikorper intraoperativ, bevorzugt direkt lokal in die Bauchhohle, verabreicht wird.

Horst Lindhofer

Papers

Induction of antitumor immunity by inducing tumor cells to antigen-presenting cells again

Procédé d'immunisation ex vivo utilisant des anticorps hétérologues intactes bispécifiques ou trispécifiques

Intraperitoneale Applikation trifunktioneller Antikörper: Ein neuartiges Konzept zur Behandlung der Peritonealkarzinose solider Tumoren

Combination of GD2-directed bispecific trifunctional antibody therapy with Pd-1 immune checkpoint blockade induces anti-neuroblastoma immunity in a syngeneic mouse model

Intraoperative trifunktionale antikörper-applikation zur prophylaxe intraperitonealer tumorzelldissemination