D'Avino, Chiara (2016) Novel approaches for breast cancer therapy: exploiting fully human immunoRNases targeting ErbB2-positive and triple-negative breast cancers. [Tesi di dottorato]

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Tipologia del documento: Tesi di dottorato
Lingua: English
Titolo: Novel approaches for breast cancer therapy: exploiting fully human immunoRNases targeting ErbB2-positive and triple-negative breast cancers
Autori:
AutoreEmail
D'Avino, Chiarachiaradavino@gmail.com
Data: 30 Marzo 2016
Numero di pagine: 51
Istituzione: Università degli Studi di Napoli Federico II
Dipartimento: Medicina Molecolare e Biotecnologie Mediche
Scuola di dottorato: Scienze Biologiche
Dottorato: Biochimica e Biologia Cellulare e Molecolare
Ciclo di dottorato: 28
Coordinatore del Corso di dottorato:
nomeemail
Arcari, Paoloarcari@unina.it
Tutor:
nomeemail
De Lorenzo, Claudia[non definito]
Data: 30 Marzo 2016
Numero di pagine: 51
Parole chiave: breast cancer; ErbB2; human monoclonal antibodies; immunotherapy; Nucleolin; phage display; triple-negative
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/10 - Biochimica
Area 05 - Scienze biologiche > BIO/11 - Biologia molecolare
Depositato il: 01 Dic 2016 08:30
Ultima modifica: 10 Mag 2017 01:00
URI: http://www.fedoa.unina.it/id/eprint/11151

Abstract

Breast cancer is the most common cancer in women worldwide. A new promising anti-cancer therapy involves the use of monoclonal antibodies specific for tumor-associated antigens (TAAs). A TAA of interest for breast cancer immunotherapy is ErbB2, a tyrosine kinase receptor overexpressed in 25-30% of breast carcinomas but not on most of normal tissues. Trastuzumab, a humanized anti-ErbB2 antibody in clinical use for the therapy of breast cancer, is effective but can cause resistance and/or cardiotoxicity. Taking advantage of phage display technology, we had previously isolated a human anti-ErbB2 scFv, named Erbicin, which recognizes an epitope different from that of Trastuzumab. We then engineered an anti-ErbB2 immunoRNase (IR), called Erb-HP-DDADD-RNase, by fusing Erbicin with a human pancreatic RNase (HP-RNase) variant, made resistant to the cytosolic RNase inhibitor (RI). Here we report that this novel IR is endowed with antiproliferative activity for Trastuzumab-resistant breast cancer cells, both in vitro and in vivo, that is more potent than that of the parental immunoRNase, which is sensitive to the RNase inhibitor. Importantly, Erb-HP-DDADD-RNase does not show cardiotoxic effects in vitro on human cardiomyocytes neither impairing cardiac function in a mouse model. Since, unfortunately, most of human breast tumors as the aggressive triple negative breast cancer (TNBC) are ErbB2-negative, we characterized a novel target for immunotherapy of TNBC to complement the promising data on IR/ErbB: Nucleolin (NCL), a multifunctional protein, selectively expressed on the surface of cancer cells, which regulates also the biogenesis of specific microRNAs (miRNAs) involved in tumor development and drug-resistance. By using phage display, we have isolated a novel human anti-NCL scFv, named 4LB5, endowed with selective anti-tumor effects. Here we report on the construction and characterization of a novel immunoRNase made up of 4LB5 and human pancreatic RNase (HP-RNase), named 4LB5-HP-RNase. This novel IR: i) retains both the enzymatic activity of human pancreatic RNase and the specific binding of the parental scFv to a panel of surface NCL-positive breast cancer cells; ii) dramatically and selectively reduces the viability and proliferation of NCL-positive tumor cells, both in vitro and in vivo; iii) induces apoptosis and strongly affects the levels of tumorigenic miRNAs, such as miR-21, -221 and 222. Altogether, these two novel immunoagents could be valuable tools for the therapeutic need of breast cancer patients ineligible for Trastuzumab treatment due to resistance, cardiotoxicity or lack of ErbB2 receptor.

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