Antibody–Drug Conjugate Efficacy in Neuroblastoma: Role of Payload, Resistance Mechanisms, Target Density, and Antibody Internalization

Buongervino, Samantha N.; Lane, Maria; Garrigan, Emily; Zhelev, Doncho V.; Dimitrov, Dimiter S.; Bosse, Kristopher R.

doi:10.1158/1535-7163.mct-20-1034

Cited by 12 publications

(17 citation statements)

References 60 publications

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“…However, multiple studies are underway exploring ADCs targeting alternative markers in GD2-expressing tumors. 1 In neuroblastoma, an ADC targeting the neural cell adhesion molecule (CD56) has recently been evaluated in the clinical setting (NCT02452554), while B7-H3, 25 anaplastic lymphoma kinase, galectin-3 binding protein, and glypican 2 26 have all been proven to be effective targets for ADCs in preclinical models. Active clinical trials explore ADCs directed to the AXL receptor tyrosine kinase in breast cancers, melanoma, and sarcomas (NCT03425279; NCT02988817), and to the B7-H3 molecule in TNBC and melanoma (NCT03729596).…”

Section: Discussionmentioning

confidence: 99%

Therapeutic efficacy of antibody-drug conjugates targeting GD2-positive tumors

Kalinovsky

Kibardin

Холоденко

et al. 2022

J Immunother Cancer

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BackgroundBoth ganglioside GD2-targeted immunotherapy and antibody-drug conjugates (ADCs) have demonstrated clinical success as solid tumor therapies in recent years, yet no research has been carried out to develop anti-GD2 ADCs against solid tumors. This is the first study to analyze cytotoxic activity of clinically relevant anti-GD2 ADCs in a wide panel of cell lines with varying GD2 expression and their effects in mouse models of GD2-positive solid cancer.MethodsAnti-GD2 ADCs were generated based on the GD2-specific antibody ch14.18 approved for the treatment of neuroblastoma and commonly used drugs monomethyl auristatin E (MMAE) or F (MMAF), conjugated via a cleavable linker by thiol-maleimide chemistry. The antibody was produced in a mammalian expression system, and its specific binding to GD2 was analyzed. Antigen-binding properties and biodistribution of the ADCs in mice were studied in comparison with the parent antibody. Cytotoxic effects of the ADCs were evaluated in a wide panel of GD2-positive and GD2-negative tumor cell lines of neuroblastoma, glioma, sarcoma, melanoma, and breast cancer. Their antitumor effects were studied in the B78-D14 melanoma and EL-4 lymphoma syngeneic mouse models.ResultsThe ch14.18-MMAE and ch14.18-MMAF ADCs retained antigen-binding properties of the parent antibody. Direct dependence of the cytotoxic effect on the level of GD2 expression was observed in cell lines of different origin for both ADCs, with IC50 below 1 nM for the cells with high GD2 expression and no cytotoxic effect for GD2-negative cells. Within the analyzed cell lines, ch14.18-MMAF was more effective in the cells overexpressing GD2, while ch14.18-MMAE had more prominent activity in the cells expressing low GD2 levels. The ADCs had a similar biodistribution profile in the B78-D14 melanoma model compared with the parent antibody, reaching 7.7% ID/g in the tumor at 48 hours postinjection. The average tumor size in groups treated with ch14.18-MMAE or ch14.18-MMAF was 2.6 times and 3.8 times smaller, respectively, compared with the control group. Antitumor effects of the anti-GD2 ADCs were also confirmed in the EL-4 lymphoma model.ConclusionThese findings validate the potential of ADCs targeting ganglioside GD2 in treating multiple GD2-expressing solid tumors.

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Section: Discussionmentioning

confidence: 99%

Therapeutic efficacy of antibody-drug conjugates targeting GD2-positive tumors

Kalinovsky

Kibardin

Холоденко

et al. 2022

J Immunother Cancer

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“…The antitumor efficacy of T-cell-engaging bispecific antibodies and antibody-drug conjugates is also dependent on antigen density on target cells and in general, a higher antigen density compared to CAR T-cells is required to unfold their therapeutic effect. 46,47 Recently, several studies have reported on dual-(or even triple-) antigen targeting with CAR T-cells against MM, e.g. with BCMA in combination with GPRC5D or SLAMF7.…”

Section: Discussionmentioning

confidence: 99%

All-trans retinoic acid works synergistically with the γ-secretase inhibitor crenigacestat to augment BCMA on multiple myeloma and the efficacy of BCMA-CAR T cells

et al. 2022

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B-cell maturation antigen (BCMA) is the lead antigen for CAR T-cell therapy in multiple myeloma (MM). A challenge is inter- and intra-patient heterogeneity in BCMA expression on MM cells and BCMA downmodulation under therapeutic pressure. Accordingly, there is a desire to augment and sustain BCMA expression on MM cells in patients that receive BCMA-CAR T-cell therapy. We used all-trans retinoic acid (ATRA) to augment BCMA expression on MM cells and to increase the efficacy of BCMA-CAR T-cells in pre-clinical models. We show that ATRA treatment leads to an increase in BCMA transcripts by quantitative PCR and an increase in BCMA protein expression by flow cytometry in MM cell lines and primary MM cells. Analyses with super-resolution microscopy confirmed increased BCMA protein expression and revealed an even distribution of non-clustered BCMA molecules on the MM cell membrane after ATRA treatment. The enhanced BCMA expression on MM cells after ATRA treatment led to enhanced cytolysis, cytokine secretion and proliferation of BCMA-CAR T-cells in vitro, and increased efficacy of BCMA-CAR T-cell therapy in a murine xenograft model of MM in vivo (NSG/MM1.S). Combination treatment of MM cells with ATRA and the γ-secretase inhibitor crenigacestat further enhanced BCMA expression and the efficacy of BCMA-CAR T-cell therapy in vitro and in vivo. Taken together, the data show that ATRA treatment leads to enhanced BCMA expression on MM cells and consecutively, enhanced reactivity of BCMA-CAR T-cells. The data support the clinical evaluation of ATRA in combination with BCMA-CAR T-cell therapy and potentially, other BCMA-directed immunotherapies.

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“…Internalization studies were performed on an Incucyte ZOOM live-cell monitoring system (Essen Bioscience) using 5 µg/mL of red-labeled D3-GPC2-IgG1 antibody, as previously detailed. 21 Cytotoxicity of D3-GPC2-PBD, AB1-SARS-CoV-2-PBD, and free PBD (Levena Biopharma) was measured using Cell Titer-Glo V.2.0 Cell Viability Assay (Promega). 21 …”

Section: Methodsmentioning

confidence: 99%

“… 21 Cytotoxicity of D3-GPC2-PBD, AB1-SARS-CoV-2-PBD, and free PBD (Levena Biopharma) was measured using Cell Titer-Glo V.2.0 Cell Viability Assay (Promega). 21 …”

Section: Methodsmentioning

confidence: 99%

GPC2 antibody–drug conjugate reprograms the neuroblastoma immune milieu to enhance macrophage-driven therapies

Pascual‐Pasto

McIntyre

Shraim

et al. 2022

J Immunother Cancer

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BackgroundAntibody–drug conjugates (ADCs) that deliver cytotoxic drugs to tumor cells have emerged as an effective and safe anticancer therapy. ADCs may induce immunogenic cell death (ICD) to promote additional endogenous antitumor immune responses. Here, we characterized the immunomodulatory properties of D3-GPC2-PBD, a pyrrolobenzodiazepine (PBD) dimer-bearing ADC that targets glypican 2 (GPC2), a cell surface oncoprotein highly differentially expressed in neuroblastoma.MethodsADC-mediated induction of ICD was studied in GPC2-expressing murine neuroblastomas in vitro and in vivo. ADC reprogramming of the neuroblastoma tumor microenvironment was profiled by RNA sequencing, cytokine arrays, cytometry by time of flight and flow cytometry. ADC efficacy was tested in combination with macrophage-driven immunoregulators in neuroblastoma syngeneic allografts and human patient-derived xenografts.ResultsThe D3-GPC2-PBD ADC induced biomarkers of ICD, including neuroblastoma cell membrane translocation of calreticulin and heat shock proteins (HSP70/90) and release of high-mobility group box 1 and ATP. Vaccination of immunocompetent mice with ADC-treated murine neuroblastoma cells promoted T cell-mediated immune responses that protected animals against tumor rechallenge. ADC treatment also reprogrammed the tumor immune microenvironment to a proinflammatory state in these syngeneic neuroblastoma models, with increased tumor trafficking of activated macrophages and T cells. In turn, macrophage or T-cell inhibition impaired ADC efficacy in vivo, which was alternatively enhanced by both CD40 agonist and CD47 antagonist antibodies. In human neuroblastomas, the D3-GPC2-PBD ADC also induced ICD and promoted tumor phagocytosis by macrophages, which was further enhanced when blocking CD47 signaling in vitro and in vivo.ConclusionsWe elucidated the immunoregulatory properties of a GPC2-targeted ADC and showed robust efficacy of combination immunotherapies in diverse neuroblastoma preclinical models.

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Antibody–Drug Conjugate Efficacy in Neuroblastoma: Role of Payload, Resistance Mechanisms, Target Density, and Antibody Internalization

Cited by 12 publications

References 60 publications

Therapeutic efficacy of antibody-drug conjugates targeting GD2-positive tumors

Therapeutic efficacy of antibody-drug conjugates targeting GD2-positive tumors

All-trans retinoic acid works synergistically with the γ-secretase inhibitor crenigacestat to augment BCMA on multiple myeloma and the efficacy of BCMA-CAR T cells

GPC2 antibody–drug conjugate reprograms the neuroblastoma immune milieu to enhance macrophage-driven therapies

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