uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
17AAG-induced internalisation of HER2-specific Affibody molecules
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Radiology, Oncology and Radiation Science, Biomedical Radiation Sciences.
Göteborgs universitet, Sahlgrenska akademin, Institutionen för kliniska vetenskaper, Avdelningen för radiofysik.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Radiology, Oncology and Radiation Science, Biomedical Radiation Sciences.
(English)Manuscript (preprint) (Other academic)
Abstract [en]

The geldanamycin derivative 17-allylamino-17-demethoxygeldanamycin (17-AAG) is known to induce internalisation and degradation of the otherwise internalisation-resistant HER2 receptor. Here we use 17-AAG to increase internalisation of the HER2-specific affibody molecule ABY-025. Cellular redistribution of 211At‑ABY025 and 111In‑ABY025 after 17-AAG treatment was studied. It was concluded that 17-AAG may be used to facilitate cell-specific intracellular localisation of a suitable cytotoxic or radioactive agent coupled to ABY-025 in HER2-overexpressing tumours.

Keyword [en]
17-AAG, internalisation, HER2, Affibody
URN: urn:nbn:se:uu:diva-156733OAI: oai:DiVA.org:uu-156733DiVA: diva2:433087
Available from: 2011-08-08 Created: 2011-08-08 Last updated: 2011-11-03
In thesis
1. Cellular Studies of HER-family Specific Affibody Molecules
Open this publication in new window or tab >>Cellular Studies of HER-family Specific Affibody Molecules
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The human epidermal growth-factor like receptor (HER) family of receptor tyrosine kinases are important targets for cancer therapy. The family consists of four members - EGFR, HER2, HER3 and HER4 - that normally transfer stimulatory signals from extracellular growth factors to the intracellular signalling network. Over-activation of these receptors leads to uncontrolled cell proliferation and is seen in several types of tumours. The aim of the studies reported in this thesis was to study the uptake and effects of affibody molecules against EGFR, HER2 and HER3 in cultured cells. Affibody molecules are affinity proteins originally derived from one of the domains of protein A, and their small size and robust structure make them suitable agents for tumour targeting and therapy.

Papers I and II of this thesis concern EGFR-specific affibody molecules, which were shown to be more similar to the antibody cetuximab than the natural ligand EGF in terms of cellular uptake, binding site and internalisation rate. In addition, fluorescence-based methods for the quantification of internalisation were evaluated.

In the studies reported in papers III and IV, HER2-specific affibody molecules were utilised as carriers of radionuclides. Paper III reports that different cell lines exhibit different radiosensitivities to 211At-labelled affibody molecules; radiosensitivity was found to correlate with cell geometry and the rate of internalisation. Paper IV discusses the use of 17-AAG, an agent that induces HER2 internalisation and degradation, to force the internalisation of 211At- and 111In-labelled affibody molecules.

Papers V and VI describe the selection and maturation of HER3-specific affibody molecules, which were found to compete with the receptor’s natural ligand, heregulin, for receptor binding. These affibody molecules were demonstrated to inhibit heregulin-induced HER3 activation and cell proliferation.

The studies summarised in this paper will hopefully contribute to a better understanding of these affibody molecules and bring them one step closer to being helpful tools in the diagnosis and treatment of cancer.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2011. 67 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 688
EGFR, HER2, HER3, Affibody, internalisation, tumour targeting
National Category
Medical and Health Sciences
urn:nbn:se:uu:diva-156730 (URN)978-91-554-8119-3 (ISBN)
Public defence
2011-09-24, Rudbecksalen, Rudbecklaboratoriet, Dag Hammarskjölds väg 20, Uppsala, 10:15 (Swedish)
Available from: 2011-09-02 Created: 2011-08-08 Last updated: 2011-11-03Bibliographically approved

Open Access in DiVA

No full text

By organisation
Biomedical Radiation Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Total: 192 hits
ReferencesLink to record
Permanent link

Direct link