nach oben

Archivum Immunologiae et Therapiae Experimentalis

Erschienen in:

01.12.2023 | Original Article

Improved Production of Anti-FGF-2 Nanobody Using Pichia pastoris and Its Effect on Antiproliferation of Keratinocytes and Alleviation of Psoriasis

verfasst von: Zhenlong Zhou, Baixin Liao, Shengli Wang, Jian Tang, Hui Zhao, Mingjie Tong, Keting Li, Sheng Xiong

Erschienen in: Archivum Immunologiae et Therapiae Experimentalis | Ausgabe 1/2023

Einloggen, um Zugang zu erhalten

Abstract

Fibroblast growth factor 2 (FGF-2) is not only an angiogenic factor, but also a mitogen for epidermal keratinocytes. FGF-2 has been shown to be positively immunoreactive in the basal layer of psoriatic lesions. In previous work, we used the Escherichia coli (E. coli) expression system to biosynthesize a biologically active anti-FGF-2 nanobody (Nb) screened by phage display technology, but the low yield limited its clinical application. In this study, we aimed to increase the yield of anti-FGF-2 Nb, and evaluate its therapeutic potential for psoriasis by inhibiting FGF-2-mediated mitogenic signaling in psoriatic epidermal keratinocytes. We demonstrated a 16-fold improvement in the yield of anti-FGF-2 Nb produced in the Pichia pastoris (P. pastoris) compared to the E. coli expression system. In vitro, the FGF-2-induced HaCaT cell model (FHCM) was established to mimic the key feature of keratinocyte overproliferation in psoriasis. Anti-FGF-2 Nb was able to effectively inhibit the proliferation and migration of FHCM. In vivo, anti-FGF-2 Nb attenuated the severity of imiquimod (IMQ)-induced psoriatic lesions in mice, and also improved the inflammatory microenvironment by inhibiting the secretion of inflammatory cytokines (IL-1β, IL-6, IL-23, and TNF-α), chemokines (CXCL1 and CCL20), and neutrophil infiltration in skin lesions. These were mainly related to the suppression of FGF-2-mediated mitogenic signaling in psoriatic keratinocytes. In conclusion, we have improved the production of anti-FGF-2 Nb and demonstrated the modality of attenuating the abnormal proliferative behavior of psoriatic keratinocytes by inhibiting FGF-2-mediated mitogenic signaling, which offers the possibility of treating psoriasis.

Vorheriger Artikel Updated Clinical Perspectives and Challenges of Chimeric Antigen Receptor-T Cell Therapy in Colorectal Cancer and Invasive Breast Cancer

Nächster Artikel Prolonged Exposure to High Glucose Induces Premature Senescence Through Oxidative Stress and Autophagy in Retinal Pigment Epithelial Cells

Ahmad M, Hirz M, Pichler H et al (2014) Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production. Appl Microbiol Biotechnol 98:5301–5317. https://doi.org/10.1007/s00253-014-5732-5CrossRefPubMedPubMedCentral

Albanesi C, Scarponi C, Giustizieri M et al (2005) Keratinocytes in inflammatory skin diseases. Curr Drug Targets Inflamm Allergy 4:329–334. https://doi.org/10.2174/1568010054022033CrossRefPubMed

Amin M, No DJ, Egeberg A et al (2018) Choosing first-line biologic treatment for moderate-to-severe psoriasis: What does the evidence say? Am J Clin Dermatol 19:1–13. https://doi.org/10.1007/s40257-017-0328-3CrossRefPubMed

Andrys C, Borska L, Pohl D et al (2007) Angiogenic activity in patients with psoriasis is significantly decreased by Goeckerman’s therapy. Arch Dermatol Res 298:479–483. https://doi.org/10.1007/s00403-006-0723-8CrossRefPubMed

Armstrong AW, Read C (2020) Pathophysiology, clinical presentation, and treatment of psoriasis: a review. JAMA 323:1945. https://doi.org/10.1001/jama.2020.4006CrossRefPubMed

Baghban R, Gargari SLM, Rajabibazl M et al (2016) Camelid-derived heavy-chain nanobody against Clostridium botulinum neurotoxin E in Pichia pastoris: Nanobody against Clostridium botulinum neurotoxin E. Biotechnol Appl Biochem 63:200–205. https://doi.org/10.1002/bab.1226CrossRefPubMed

Battaglin F, Puccini A, Ahcene Djaballah S et al (2019) The impact of panitumumab treatment on survival and quality of life in patients with RAS wild-type metastatic colorectal cancer. Cancer Manag Res 11:5911–5924. https://doi.org/10.2147/CMAR.S186042CrossRefPubMedPubMedCentral

Beatty JD, Beatty BG, Vlahos WG (1987) Measurement of monoclonal antibody affinity by non-competitive enzyme immunoassay. J Immunol Methods 100:173–179. https://doi.org/10.1016/0022-1759(87)90187-6CrossRefPubMed

Chen C-H, Poucher S, Lu J et al (2004) Fibroblast growth factor 2: from laboratory evidence to clinical application. Curr Vasc Pharmacol 2:33–43. https://doi.org/10.2174/1570161043476500CrossRefPubMed

Cos O, Ramón R, Montesinos JL et al (2006) Operational strategies, monitoring and control of heterologous protein production in the methylotrophic yeast Pichia pastoris under different promoters: a review. Microb Cell Fact 5:17. https://doi.org/10.1186/1475-2859-5-17CrossRefPubMedPubMedCentral

Dailey L, Ambrosetti D, Mansukhani A et al (2005) Mechanisms underlying differential responses to FGF signaling. Cytokine Growth Factor Rev 16:233–247. https://doi.org/10.1016/j.cytogfr.2005.01.007CrossRefPubMed

Ferrara N (1999) Role of vascular endothelial growth factor in the regulation of angiogenesis. Kidney Int 56:794–814. https://doi.org/10.1046/j.1523-1755.1999.00610.xCrossRefPubMed

Gao J, Chen F, Fang H et al (2020) Daphnetin inhibits proliferation and inflammatory response in human HaCaT keratinocytes and ameliorates imiquimod-induced psoriasis-like skin lesion in mice. Biol Res 53:48. https://doi.org/10.1186/s40659-020-00316-0CrossRefPubMedPubMedCentral

Garzorz-Stark N, Eyerich K (2019) Psoriasis pathogenesis: keratinocytes are back in the spotlight. J Invest Dermatol 139:995–996. https://doi.org/10.1016/j.jid.2019.01.026CrossRefPubMed

Han LK, Oanh NCK, Nghia NH et al (2020) Secretory expression of the recombinant FGF-2 protein in Pichia pastoris carrying multiple copies of target gene. VNUHCM J Sci Technol Dev 23:499–507. https://doi.org/10.32508/stdj.v23i2.1746CrossRef

Harper EG, Guo C, Rizzo H et al (2009) Th17 cytokines stimulate CCL20 expression in keratinocytes in vitro and in vivo: implications for psoriasis pathogenesis. J Invest Dermatol 129:2175–2183. https://doi.org/10.1038/jid.2009.65CrossRefPubMedPubMedCentral

Haugh I, Preston A, Kivelevitch D et al (2018) Risankizumab: an anti-IL-23 antibody for the treatment of psoriasis. Drug Des Devel Ther 12:3879–3883. https://doi.org/10.2147/DDDT.S167149CrossRefPubMedPubMedCentral

Hoshida H, Fujita T, Cha-aim K et al (2013) N-glycosylation deficiency enhanced heterologous production of a Bacillus licheniformis thermostable α-amylase in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 97:5473–5482. https://doi.org/10.1007/s00253-012-4582-2CrossRefPubMed

Hu Y, Liu C, Muyldermans S (2017) Nanobody-based delivery systems for diagnosis and targeted tumor therapy. Front Immunol 8:1442. https://doi.org/10.3389/fimmu.2017.01442CrossRefPubMedPubMedCentral

Jabeen M, Boisgard A-S, Danoy A et al (2020) Advanced characterization of imiquimod-induced psoriasis-like mouse model. Pharmaceutics 12:789. https://doi.org/10.3390/pharmaceutics12090789CrossRefPubMedPubMedCentral

Jiang B-W, Zhang W-J, Wang Y et al (2020) Convallatoxin induces HaCaT cell necroptosis and ameliorates skin lesions in psoriasis-like mouse models. Biomed Pharmacother 121:109615. https://doi.org/10.1016/j.biopha.2019.109615CrossRefPubMed

Jovčevska I, Muyldermans S (2020) The therapeutic potential of nanobodies. BioDrugs 34:11–26. https://doi.org/10.1007/s40259-019-00392-zCrossRefPubMed

Kijanka M, Dorresteijn B, Oliveira S et al (2015) Nanobody-based cancer therapy of solid tumors. Nanomedicine 10:161–174. https://doi.org/10.2217/nnm.14.178CrossRefPubMed

Kormeili T, Lowe NJ, Yamauchi PS (2004) Psoriasis: immunopathogenesis and evolving immunomodulators and systemic therapies; U.S. experiences. Br J Dermatol 151:3–15. https://doi.org/10.1111/j.1365-2133.2004.06009.xCrossRefPubMed

Kovács N, Farsang R, Szigeti M et al (2022) Enhanced recombinant protein production of soluble, highly active and immobilizable PNGase F. Mol Biotechnol 64:914–918. https://doi.org/10.1007/s12033-022-00464-6CrossRefPubMedPubMedCentral

Kuang Y-H, Lu Y, Liu Y-K et al (2018) Topical Sunitinib ointment alleviates Psoriasis-like inflammation by inhibiting the proliferation and apoptosis of keratinocytes. Eur J Pharmacol 824:57–63. https://doi.org/10.1016/j.ejphar.2018.01.048CrossRefPubMed

Landeck L, Sabat R, Ghoreschi K et al (2021) Immunotherapy in psoriasis. Immunotherapy 13:605–619. https://doi.org/10.2217/imt-2020-0292CrossRefPubMed

Ledsgaard L, Ljungars A, Rimbault C et al (2022) Advances in antibody phage display technology. Drug Discov Today 27:2151–2169. https://doi.org/10.1016/j.drudis.2022.05.002CrossRefPubMed

Li H, Yao Q, Mariscal AG et al (2018) Epigenetic control of IL-23 expression in keratinocytes is important for chronic skin inflammation. Nat Commun 9:1420. https://doi.org/10.1038/s41467-018-03704-zCrossRefPubMedPubMedCentral

Liu M, Li L, Jin D et al (2021) Nanobody—a versatile tool for cancer diagnosis and therapeutics. Wiley Interdiscip Rev Nanomed Nanobiotechnol 13:e1697. https://doi.org/10.1002/wnan.1697CrossRefPubMed

Moos S, Mohebiany AN, Waisman A et al (2019) Imiquimod-induced psoriasis in mice depends on the IL-17 signaling of keratinocytes. J Invest Dermatol 139:1110–1117. https://doi.org/10.1016/j.jid.2019.01.006CrossRefPubMed

Paz-Ares L, Socinski MA, Shahidi J et al (2016) Correlation of EGFR-expression with safety and efficacy outcomes in SQUIRE: a randomized, multicenter, open-label, phase III study of gemcitabine–cisplatin plus necitumumab versus gemcitabine–cisplatin alone in the first-line treatment of patients with stage IV squamous non-small-cell lung cancer. Ann Oncol 27:1573–1579. https://doi.org/10.1093/annonc/mdw214CrossRefPubMedPubMedCentral

Peng A, Gaitonde P, Kosloski MP et al (2009) Effect of route of administration of human recombinant factor VIII on its immunogenicity in hemophilia a mice. J Pharm Sci 98:4480–4484. https://doi.org/10.1002/jps.21765CrossRefPubMedPubMedCentral

Salvador J-P, Vilaplana L, Marco M-P (2019) Nanobody: outstanding features for diagnostic and therapeutic applications. Anal Bioanal Chem 411:1703–1713. https://doi.org/10.1007/s00216-019-01633-4CrossRefPubMed

Shi H, Zhou H, Ma A et al (2019) Oxymatrine therapy inhibited epidermal cell proliferation and apoptosis in severe plaque psoriasis. Br J Dermatol 181:1028–1037. https://doi.org/10.1111/bjd.17852CrossRefPubMed

Song H, Pan B, Yi J et al (2014) Featured article: autophagic activation with Nimotuzumab enhanced chemosensitivity and radiosensitivity of esophageal squamous cell carcinoma. Exp Biol Med 239:529–541. https://doi.org/10.1177/1535370214525315CrossRef

Sun X, Zhang C, Jin H et al (2016) Flow cytometric analysis of T lymphocyte proliferation in vivo by EdU incorporation. Int Immunopharmacol 41:56–65. https://doi.org/10.1016/j.intimp.2016.10.019CrossRefPubMed

Svecova D, Lubell MW, Casset-Semanaz F et al (2019) A randomized, double-blind, placebo-controlled phase 1 study of multiple ascending doses of subcutaneous M1095, an anti-interleukin 17A/F nanobody, in moderate-to-severe psoriasis. J Am Acad Dermatol 81:196–203. https://doi.org/10.1016/j.jaad.2019.03.056CrossRefPubMed

Tang J, Liu C, Liu S et al (2022a) Inhibition of JAK1/STAT3 pathway by 2-methoxyestradiol ameliorates psoriatic features in vitro and in an imiquimod-induced psoriasis-like mouse model. Eur J Pharmacol 933:175276. https://doi.org/10.1016/j.ejphar.2022.175276CrossRefPubMed

Tang J, Zhou Z, Qian J et al (2022b) KdPT alleviates imiquimod-induced psoriasis-like skin lesion in mice via inhibiting proliferation and inflammation response. Pharmazie 77:48–53PubMed

Tol J, Punt CJA (2010) Monoclonal antibodies in the treatment of metastatic colorectal cancer: a review. Clin Ther 32:437–453. https://doi.org/10.1016/j.clinthera.2010.03.012CrossRefPubMed

Van Belle AB, de Heusch M, Lemaire MM et al (2012) IL-22 is required for imiquimod-induced psoriasiform skin inflammation in mice. J Immunol 188:462–469. https://doi.org/10.4049/jimmunol.1102224CrossRefPubMed

van der Fits L, Mourits S, Voerman JSA et al (2009) Imiquimod-induced psoriasis-like skin inflammation in mice is mediated via the IL-23/IL-17 axis. J Immunol 182:5836–5845. https://doi.org/10.4049/jimmunol.0802999CrossRefPubMed

Vivanco I, Mellinghoff IK (2010) Epidermal growth factor receptor inhibitors in oncology. Curr Opin Oncol 22:573–578. https://doi.org/10.1097/CCO.0b013e32833edbdfCrossRefPubMed

Wang P, Zhang J, Sun Z et al (2000) Glycosylation of prourokinase produced by Pichia pastoris impairs enzymatic activity but not secretion. Protein Expr Purif 20:179–185. https://doi.org/10.1006/prep.2000.1310CrossRefPubMed

Wang L, Zhang G, Qin L et al (2020) Anti-EGFR binding nanobody delivery system to improve the diagnosis and treatment of solid tumours. Recent Pat Anticancer Drug Discov 15:200–211. https://doi.org/10.2174/1574892815666200904111728CrossRefPubMed

Werner S, Smola H (2001) Paracrine regulation of keratinocyte proliferation and differentiation. Trends Cell Biol 11:143–146. https://doi.org/10.1016/S0962-8924(01)01955-9CrossRefPubMed

Wu X, Huang H, Wang C et al (2013) Identification of a novel peptide that blocks basic fibroblast growth factor-mediated cell proliferation. Oncotarget 4:1819–1828. https://doi.org/10.18632/oncotarget.1312CrossRefPubMedPubMedCentral

Xiao Y, Wang C, Zeng B et al (2020) miR124-3p/FGFR2 axis inhibits human keratinocyte proliferation and migration and improve the inflammatory microenvironment in psoriasis. Mol Immunol 122:89–98. https://doi.org/10.1016/j.molimm.2020.04.002CrossRefPubMed

Xiong S, Xie Q, Chen C et al (2022) Application of anti-recombinant human basic fibroblast growth factor nanobody. CN Patent CN113150141 B

Xu Y, Zhu J, Hu J et al (2021) L-theanine alleviates IMQ-induced psoriasis like skin inflammation by downregulating the production of IL-23 and chemokines. Front Pharmacol 12:719842. https://doi.org/10.3389/fphar.2021.719842CrossRefPubMedPubMedCentral

Yaguchi H, Tsuboi R, Ueki R et al (1993) Immunohistochemical localization of basic fibroblast growth factor in skin diseases. Acta Derm Venereol 73:81–83CrossRefPubMed

Yang EY, Shah K (2020) Nanobodies: next generation of cancer diagnostics and therapeutics. Front Oncol 10:1182. https://doi.org/10.3389/fonc.2020.01182CrossRefPubMedPubMedCentral

Zhang C, Zhang W, Tang X et al (2020) Change of amino acid residues in idiotypic nanobodies enhanced the sensitivity of competitive enzyme immunoassay for mycotoxin ochratoxin A in Cereals. Toxins 12:273. https://doi.org/10.3390/toxins12040273CrossRefPubMedPubMedCentral

Zhou X, Chen Y, Cui L et al (2022) Advances in the pathogenesis of psoriasis: from keratinocyte perspective. Cell Death Dis 13:81. https://doi.org/10.1038/s41419-022-04523-3CrossRefPubMedPubMedCentral

Zittermann SI, Issekutz AC (2006) Basic fibroblast growth factor (bFGF, FGF-2) potentiates leukocyte recruitment to inflammation by enhancing endothelial adhesion molecule expression. Am J Pathol 168:835–846. https://doi.org/10.2353/ajpath.2006.050479CrossRefPubMedPubMedCentral

Titel: Improved Production of Anti-FGF-2 Nanobody Using Pichia pastoris and Its Effect on Antiproliferation of Keratinocytes and Alleviation of Psoriasis
verfasst von: Zhenlong Zhou
Baixin Liao
Shengli Wang
Jian Tang
Hui Zhao
Mingjie Tong
Keting Li
Sheng Xiong
Publikationsdatum: 01.12.2023
Verlag: Springer International Publishing
Erschienen in: Archivum Immunologiae et Therapiae Experimentalis / Ausgabe 1/2023
Print ISSN: 0004-069X
Elektronische ISSN: 1661-4917
DOI: https://doi.org/10.1007/s00005-023-00685-w

Springer Medizin Österreich

Abstract

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 1/2023

Updated Clinical Perspectives and Challenges of Chimeric Antigen Receptor-T Cell Therapy in Colorectal Cancer and Invasive Breast Cancer

Stress-Sensitive Regulators of Fetal Neurodevelopment in HIV and Preeclampsia: An Immunocytochemical Appraisal of Placental OGT and T4 Levels

Modulation of Myeloid-Derived Suppressor Cells in the Tumor Microenvironment by Natural Products

Activating NKG2C Receptor: Functional Characteristics and Current Strategies in Clinical Applications

Investigation of the Immunogenic Properties of Ovalbumin Modified by Urban Airborne Particulate Matter

The Pyroptotic and Nonpyroptotic Roles of Gasdermins in Modulating Cancer Progression and Their Perspectives on Cancer Therapeutics