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

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

doi:10.3892/ol.2021.13120

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

Authors:
- Luigi Grasso
- James Bradford Kline
- Nicholas C. Nicolaides
View Affiliations

Affiliations: Navrogen Inc., Cheyney, PA 19319, USA
Published online on: November 2, 2021 https://doi.org/10.3892/ol.2021.13120
Article Number: 2
Copyright: © Grasso et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Rituximab (RTX) is a CD20‑targeting antibody that is the standard‑of‑care for patients with non‑Hodgkin Lymphoma (NHL) cases. RTX's mechanism of action includes complement‑dependent cytotoxicity (CDC) and antibody‑dependent cellular cytotoxicity (ADCC). Recent clinical evidence suggests that high serum levels of the tumor‑produced mucin 16 (MUC16) and cancer antigen 125 (CA125) have a negative impact on the effectiveness of RTX clinical activity in up to 40% of patients with follicular lymphoma. The present study sought to understand the possible mechanism underlying these results; therefore, cellular and molecular analyses of RTX and CA125 interaction were peformed, and a library of RTX variants was generated using a proprietary technology called Block‑Removed Immunoglobulin Technology that combines randomized amino acid substitutions and high‑throughput functional screenings to identify CA125‑refractory RTX variants. The present study demonstrated that CA125 could bind to RTX and reduce its tumor cell killing activity. Furthermore, the study characterized an RTX variant, named NAV‑006 (RTX‑N109D), which was more refractory to the immunosuppressive effects mediated by CA125 as evidenced by its reduced CA125 interaction and increased activity of ADCC and CDC when compared with parent RTX. Taken together, these findings warranted further investigation on NAV‑006 as a next generation anti‑CD20 antibody that could improve the efficacy of parent RTX in NHL patients with high levels of CA125.

View Figures

View References

1	National Cancer Institute, . Surveillance, Epidemiology, and End Results (SEER) Program: Cancer Stat Facts: NHL — Follicular Lymphoma. https://seer.cancer.gov/statfacts/html/follicular.htmlOctober 27–2021
2	Gallagher CJ, Gregory WM, Jones AE, Stansfeld AG, Richards MA, Dhaliwal HS, Malpas JS and Lister TA: Follicular lymphoma: Prognostic factors for response and survival. J Clin Oncol. 4:1470–1480. 1986. View Article : Google Scholar : PubMed/NCBI
3	Manches O, Lui G, Chaperot L, Gressin R, Molens JP, Jacob MC, Sotto JJ, Leroux D, Bensa JC and Plumas J: In vitro mechanisms of action of rituximab on primary non-Hodgkin lymphomas. Blood. 101:949–954. 2003. View Article : Google Scholar : PubMed/NCBI
4	Clynes RA, Towers TL, Presta LG and Ravetch JV: Inhibitory Fc receptors modulate in vivo cytotoxicity against tumor targets. Nat Med. 6:443–446. 2000. View Article : Google Scholar : PubMed/NCBI
5	Cartron G, Dacheux L, Salles G, Solal-Celigny P, Bardos P, Colombat P and Watier H: Therapeutic activity of humanized anti-CD20 monoclonal antibody and polymorphism in IgG Fc receptor FcgammaRIIIa gene. Blood. 99:754–758. 2002. View Article : Google Scholar : PubMed/NCBI
6	Seyfizadeh N, Seyfizadeh N, Hasenkamp J and Huerta-Yepez S: A molecular perspective on rituximab: A monoclonal antibody for B cell non Hodgkin lymphoma and other affections. Crit Rev Oncol Hematol. 97:275–290. 2016. View Article : Google Scholar : PubMed/NCBI
7	Scott DW and Gascoyne RD: The tumour microenvironment in B cell lymphomas. Nat Rev Cancer. 14:517–534. 2014. View Article : Google Scholar : PubMed/NCBI
8	Dilek I, Ayakta H, Demir C, Meral C and Ozturk M: CA 125 levels in patients with non-Hodgkin lymphoma and other hematologic malignancies. Clin Lab Haematol. 27:51–55. 2005. View Article : Google Scholar : PubMed/NCBI
9	Fehm T, Beck E, Valerius T, Gramatzki M and Jäger W: CA 125 elevations in patients with malignant lymphomas. Tumour Biol. 19:283–289. 1998. View Article : Google Scholar : PubMed/NCBI
10	Lazzarino M, Orlandi E, Klersy C, Astori C, Brusamolino E, Corso A, Bellio L, Gargantini L, Morra E and Bernasconi C: Serum CA 125 is of clinical value in the staging and follow-up of patients with non-Hodgkin's lymphoma: Correlation with tumor parameters and disease activity. Cancer. 82:576–582. 1998. View Article : Google Scholar : PubMed/NCBI
11	Gutiérrez A, Martínez-Serra J, Barceló B, Sampol A, Viñas L, González G, Bea MD, Amat JC, Martín J, Ramos R, et al: Prognostic value of serum CA125 levels in diffuse large B-cell lymphoma: Potential role of a new sex- and age-adjusted reference value. Int J Lab Hematol. 32:582–589. 2010. View Article : Google Scholar : PubMed/NCBI
12	Memar B, Aledavood A, Shahidsales S, Ahadi M, Farzadnia M, Raziee HR, Noori S, Tayebi-Meybodi N, Amouian S and Mohtashami S: The prognostic role of tumor marker CA-125 in B-cell non-Hodgkin's lymphoma. Iran J Cancer Prev. 8:42–46. 2015.PubMed/NCBI
13	Kline JB, Kennedy RP, Albone E, Chao Q, Fernando S, McDonough JM, Rybinski K, Wang W, Somers EB, Schweizer C, et al: Tumor antigen CA125 suppresses antibody-dependent cellular cytotoxicity (ADCC) via direct antibody binding and suppressed Fc-γ receptor engagement. Oncotarget. 8:52045–52060. 2017. View Article : Google Scholar : PubMed/NCBI
14	Vergote I, Armstrong D, Scambia G, Teneriello M, Sehouli J, Schweizer C, Weil SC, Bamias A, Fujiwara K, Ochiai K, et al: A randomized, double-blind, placebo-controlled, phase III study to assess efficacy and safety of weekly farletuzumab in combination with carboplatin and taxane in patients with ovarian cancer in first platinum-sensitive relapse. J Clin Oncol. 34:2271–2278. 2016. View Article : Google Scholar : PubMed/NCBI
15	Nicolaides NC, Schweizer C, Somers EB, Wang W, Fernando S, Ross EN, Grasso L, Hassan R and Kline JB: CA125 suppresses amatuximab immune-effector function and elevated serum levels are associated with reduced clinical response in first line mesothelioma patients. Cancer Biol Ther. 19:622–630. 2018. View Article : Google Scholar : PubMed/NCBI
16	Lin J, Spidel JL, Maddage CJ, Rybinski KA, Kennedy RP, Krauthauser CL, Park YC, Albone EF, Jacob S, Goserud MT, et al: The antitumor activity of the human FOLR1-specific monoclonal antibody, farletuzumab, in an ovarian cancer mouse model is mediated by antibody-dependent cellular cytotoxicity. Cancer Biol Ther. 14:1032–1038. 2013. View Article : Google Scholar : PubMed/NCBI
17	Ebel W, Routhier EL, Foley B, Jacob S, McDonough JM, Patel RK, Turchin HA, Chao Q, Kline JB, Old LJ, et al: Preclinical evaluation of MORAb-003, a humanized monoclonal antibody antagonizing folate receptor-alpha. Cancer Immun. 7:62007.PubMed/NCBI
18	Hassan R, Ebel W, Routhier EL, Patel R, Kline JB, Zhang J, Chao Q, Jacob S, Turchin H, Gibbs L, et al: Preclinical evaluation of MORAb-009, a chimeric antibody targeting tumor-associated mesothelin. Cancer Immun. 7:202007.PubMed/NCBI
19	Kline JB, Fernando S, Ross EN, Grasso L and Nicolaides NC: Tumor-shed antigen CA125 blocks complement-mediated killing via suppression of C1q-antibody binding. Eur J Immunol. 48:1872–1882. 2018. View Article : Google Scholar : PubMed/NCBI
20	Weng WK and Levy R: Two immunoglobulin G fragment C receptor polymorphisms independently predict response to rituximab in patients with follicular lymphoma. J Clin Oncol. 21:3940–3947. 2003. View Article : Google Scholar : PubMed/NCBI
21	Hatjiharissi E, Xu L, Santos DD, Hunter ZR, Ciccarelli BT, Verselis S, Modica M, Cao Y, Manning RJ, Leleu X, et al: Increased natural killer cell expression of CD16, augmented binding and ADCC activity to rituximab among individuals expressing the Fc{gamma}RIIIa-158 V/V and V/F polymorphism. Blood. 110:2561–2564. 2007. View Article : Google Scholar : PubMed/NCBI
22	Procházka V, Faber E, Raida L, Kapitáňová Z, Langová K, Indrák K and Papajík T: High serum carbohydrate antigen-125 (CA-125) level predicts poor outcome in patients with follicular lymphoma independently of the FLIPI score. Int J Hematol. 96:58–64. 2012. View Article : Google Scholar : PubMed/NCBI
23	Painter RH, Foster DB, Gardner B and Hughes-Jones NC: Functional affinity constants of subfragments of immunoglobulin G for Clq. Mol Immunol. 19:127–131. 1982. View Article : Google Scholar : PubMed/NCBI
24	Chappel MS, Isenman DE, Everett M, Xu YY, Dorrington KJ and Klein MH: Identification of the Fc gamma receptor class I binding site in human IgG through the use of recombinant IgG1/IgG2 hybrid and point-mutated antibodies. Proc Natl Acad Sci USA. 88:9036–9040. 1991. View Article : Google Scholar : PubMed/NCBI
25	Orlandi C, Deredge D, Ray K, Gohain N, Tolbert W, DeVico AL, Wintrode P, Pazgier M and Lewis GK: Antigen-induced allosteric changes in a human IgG1 Fc increase low-affinity Fcγ receptor binding. Structure. 28:516–527.e5. 2020. View Article : Google Scholar : PubMed/NCBI
26	Dam TK, Torres M, Brewer CF and Casadevall A: Isothermal titration calorimetry reveals differential binding thermodynamics of variable region-identical antibodies differing in constant region for a univalent ligand. J Biol Chem. 283:31366–31370. 2008. View Article : Google Scholar : PubMed/NCBI
27	Pritsch O, Magnac C, Dumas G, Bouvet JP, Alzari P and Dighiero G: Can isotype switch modulate antigen-binding affinity and influence clonal selection? Eur J Immunol. 30:3387–3395. 2000. View Article : Google Scholar : PubMed/NCBI
28	Regep C, Georges G, Shi J, Popovic B and Deane CM: The H3 loop of antibodies shows unique structural characteristics. Proteins. 85:1311–1318. 2017. View Article : Google Scholar : PubMed/NCBI
29	Rougé L, Chiang N, Steffek M, Kugel C, Croll TI, Tam C, Estevez A, Arthur CP, Koth CM, Ciferri C, et al: Structure of CD20 in complex with the therapeutic monoclonal antibody rituximab. Science. 367:1224–1230. 2020. View Article : Google Scholar : PubMed/NCBI
30	Denoel A, Dieude P, Chollet-Martin S and Grootenboer-Mignot S: SAT0181 immunogenicity of rituximab in patients with rheumatoid arthritis: A kinetic analysis. Ann Rheum Dis. 74 (Suppl≈2):S720.2–720. 2015. View Article : Google Scholar
31	Dhanda SK, Karosiene E, Edwards L, Grifoni A, Paul S, Andreatta M, Weiskopf D, Sidney J, Nielsen M, Peters B and Sette A: Predicting HLA CD4 immunogenicity in human populations. Front Immunol. 9:13692018. View Article : Google Scholar : PubMed/NCBI