Block‑Removed Immunoglobulin Technology to enhance rituximab effector function by counteracting CA125‑mediated immunosuppression

Grasso, Luigi; Kline, J. Bradford; Nicolaides, Nicholas C.

doi:10.3892/ol.2021.13120

Cited by 2 publications

(10 citation statements)

References 29 publications

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“…This has been highlighted by studies monitoring the effects of HIO factors such as MUC16/CA125 and MUC1 on humoral immune effector activities against tumor cells. The direct binding of the immunosuppressive MUC16/CA125 to IgG1-type antibodies has been shown to suppress immune effector activities and is associated with decreased therapeutic activity in human clinical studies [5][6][7]. MUC1 has been found to be overexpressed by several carcinomas and its presence has been associated with immunosuppression of ADCC; however, its mechanism of immune effector suppression has not been elucidated [18].…”

Section: Nav-003 Activity Against Humoral Immunosuppressive Tumor Cel...mentioning

confidence: 99%

“…After selection, pools were subcloned by limiting dilution and conditioned media were tested for recombinant NAV-003 antibody concentrations via ELISA using an anti-human IgG1 capture and anti-human Fc-HRP probe as described below. The best production clones were expanded for NAV-003 isolation via protein A chromatography as previously described [6] and analyzed for homogeneity via SDS-PAGE and antigen binding via ELISA and/or immunofluorescent cell staining as described below. To generate deglycosylated NAV-003 (DeGly NAV-003), 10 mg of NAV-003 was deglycosylated enzymatically using GlycINA-TOR MidiSpin column following the manufacturer's instructions (Genovis Inc., Cambridge, MA).…”

Section: Proteins Antibodies and Bispecific Antibodymentioning

confidence: 99%

“…ELISA and immunofluorescent cell staining assays were used to measure MSLN, CD3ε, IL-2, and human serum albumin (HSA, negative control) binding of antibodies as previously described [5,6]. Briefly, for testing of anti-MSLN and anti-CD3ε binding, clear 96-well plates were coated with 100 ng/mL of recombinant MSLN (Sino Biological), CD3ε (Sino Biological), or 100 ng/mL of HSA (Sigma) in 0.05 M carbonate buffer, pH 9.5 overnight at 4°C.…”

Section: Elisa and Immunofluorescence Cell Stainingmentioning

confidence: 99%

“…Correspondence: Dr. Luigi Grasso e-mail: luigi@navrogen.com While the function of MSLN is not completely understood, one defined role has been its involvement in heterotypic cellular adhesion where it has been found to bind the MUC16/CA125 protein on neighboring cells for cellular homeostasis and migration [2,3]. While several anti-MSLN-targeted approaches are being pursued to treat MSLN-positive cancers [4], a new focus has been the development of antibodies that are refractory to immunosuppressive tumor produced proteins, referred to as humoral immuno-oncology (HIO) factors, which can suppress immune effector activities, or antibodies that can target epitopes optimal for cytotoxic immune effector activity [5,6]. Previous studies have found that HIO factors such as MUC16/CA125 can bind to subsets of IgG1 antibodies and suppress their immune effector activities, including antibody-dependent cellular cytotoxicity (ADCC) and/or complement-dependent cytotoxicity (CDC) via blockade of CD16a Fc-gamma receptor and C1q protein IgG1 Fc binding, respectively [5][6][7].…”

Section: Introductionmentioning

confidence: 99%

“…While several anti-MSLN-targeted approaches are being pursued to treat MSLN-positive cancers [4], a new focus has been the development of antibodies that are refractory to immunosuppressive tumor produced proteins, referred to as humoral immuno-oncology (HIO) factors, which can suppress immune effector activities, or antibodies that can target epitopes optimal for cytotoxic immune effector activity [5,6]. Previous studies have found that HIO factors such as MUC16/CA125 can bind to subsets of IgG1 antibodies and suppress their immune effector activities, including antibody-dependent cellular cytotoxicity (ADCC) and/or complement-dependent cytotoxicity (CDC) via blockade of CD16a Fc-gamma receptor and C1q protein IgG1 Fc binding, respectively [5][6][7]. Recently, we have discovered a new HIO factor, HIO-3, that binds to the CH3 domain of IgG1-type antibodies and suppresses the immune effector activity of tumortargeting antibodies by reducing antibody-CD16a and antibody-C1q binding.…”

Section: Introductionmentioning

confidence: 99%

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NAV‐003, a bispecific antibody targeting a unique mesothelin epitope and CD3ε with improved cytotoxicity against humoral immunosuppressed tumors

Grasso¹,

Jiang

Hassan

et al. 2023

Eur J Immunol

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Mesothelin (MSLN) is a cell surface protein overexpressed in a number of cancer types. Several antibody‐ and cellular‐based MSLN targeting agents have been tested in clinical trials where their therapeutic efficacy has been moderate at best. Previous studies using antibody and Chimeric Antigen Receptor–T cells (CAR‐T) strategies have shown the importance of particular MSLN epitopes for optimal therapeutic response, while other studies have found that certain MSLN‐positive tumors can produce proteins that can bind to subsets of IgG1‐type antibodies and suppress their immune effector activities. In an attempt to develop an improved anti‐MSLN targeting agent, we engineered a humanized divalent anti‐MSLN/anti‐CD3ε bispecific antibody that avoids suppressive factors, can target a MSLN epitope proximal to the tumor cell surface, and is capable of effectively binding, activating, and redirecting T cells to the surface of MSLN‐positive tumor cells. NAV‐003 has shown significantly improved tumor cell killing against lines producing immunosuppressive proteins in vitro and in vivo. Moreover, NAV‐003 demonstrated good tolerability in mice and efficacy against patient‐derived mesothelioma xenografts co‐engrafted with human peripheral blood mononuclear cells. Together these data support the potential for NAV‐003 clinical development and human proof‐of‐concept studies in patients with MSLN‐expressing cancers.

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Section: Nav-003 Activity Against Humoral Immunosuppressive Tumor Cel...mentioning

confidence: 99%

Section: Proteins Antibodies and Bispecific Antibodymentioning

confidence: 99%

Section: Elisa and Immunofluorescence Cell Stainingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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NAV‐003, a bispecific antibody targeting a unique mesothelin epitope and CD3ε with improved cytotoxicity against humoral immunosuppressed tumors

Grasso¹,

Jiang

Hassan

et al. 2023

Eur J Immunol

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NAV-001, a high-efficacy antibody-drug conjugate targeting mesothelin with improved delivery of a potent payload by counteracting MUC16/CA125 inhibitory effects

2023

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Subsets of tumor-produced cell surface and secreted proteins can bind to IgG1 type antibodies and suppress their immune-effector activities. As they affect antibody and complement-mediated immunity, we call these proteins humoral immuno-oncology (HIO) factors. Antibody-drug conjugates (ADCs) use antibody targeting to bind cell surface antigens, internalize into the cell, then kill target cells upon liberation of the cytotoxic payload. Binding of the ADC antibody component by a HIO factor may potentially hamper ADC efficacy due to reduced internalization. To determine the potential effects of HIO factor ADC suppression, we evaluated the efficacy of a HIO-refractory, mesothelin-directed ADC (NAV-001) and a HIO-bound, mesothelin-directed ADC (SS1). The HIO factor MUC16/CA125 binding to SS1 ADC was shown to have a negative effect on internalization and tumor cell killing. The MUC16/CA125 refractory NAV-001 ADC was shown to have robust killing of MUC16/CA125 expressing and non-expressing tumor cells in vitro and in vivo at single, sub-mg/kg dosing. Moreover, NAV-001-PNU, which contains the PNU-159682 topoisomerase II inhibitor, demonstrated good stability in vitro and in vivo as well as robust bystander activity of resident cells while maintaining a tolerable safety profile in vivo. Single-dose NAV-001-PNU demonstrated robust tumor regression of a number of patient-derived xenografts from different tumor types regardless of MUC16/CA125 expression. These findings suggest that identification of HIO-refractory antibodies to be used in ADC format may improve therapeutic efficacy as observed by NAV-001 and warrants NAV-001-PNU’s advancement to human clinical trials as a monotherapy to treat mesothelin-positive cancers.

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Block‑Removed Immunoglobulin Technology to enhance rituximab effector function by counteracting CA125‑mediated immunosuppression

Cited by 2 publications

References 29 publications

NAV‐003, a bispecific antibody targeting a unique mesothelin epitope and CD3ε with improved cytotoxicity against humoral immunosuppressed tumors

NAV‐003, a bispecific antibody targeting a unique mesothelin epitope and CD3ε with improved cytotoxicity against humoral immunosuppressed tumors

NAV-001, a high-efficacy antibody-drug conjugate targeting mesothelin with improved delivery of a potent payload by counteracting MUC16/CA125 inhibitory effects

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