EGFR endocytosis requires its kinase activity and N-terminal transmembrane dimerization motif

Heukers, Raimond; Vermeulen, Jeroen F.; Fereidouni, Farzad; Bader, Arjen N.; Voortman, Jarno; Roovers, Rob C.; Gerritsen, Hans C.; Henegouwen, Paul M.P. van Bergen en

doi:10.1242/jcs.128611

Cited by 54 publications

(63 citation statements)

References 75 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Receptor dimerization stimulates EGFR endocytosis for turnover and downregulation of EGFRs4647. Anti-EGFR monoclonal antibodies, such as cetuximab and necitumumab, recognize the domain III ligand-binding region of EGFR and rapidly trigger crosslinking of EGFRs due to their bivalent format, thereby stimulating internalization and degradation of the receptor48.…”

Section: Discussionmentioning

confidence: 99%

Conditional internalization of PEGylated nanomedicines by PEG engagers for triple negative breast cancer therapy

Burnouf

Chuang

et al. 2017

Nat Commun

View full text Add to dashboard Cite

Triple-negative breast cancer (TNBC) lacks effective treatment options due to the absence of traditional therapeutic targets. The epidermal growth factor receptor (EGFR) has emerged as a promising target for TNBC therapy because it is overexpressed in about 50% of TNBC patients. Here we describe a PEG engager that simultaneously binds polyethylene glycol and EGFR to deliver PEGylated nanomedicines to EGFR+ TNBC. The PEG engager displays conditional internalization by remaining on the surface of TNBC cells until contact with PEGylated nanocarriers triggers rapid engulfment of nanocargos. PEG engager enhances the anti-proliferative activity of PEG-liposomal doxorubicin to EGFR+ TNBC cells by up to 100-fold with potency dependent on EGFR expression levels. The PEG engager significantly increases retention of fluorescent PEG probes and enhances the antitumour activity of PEGylated liposomal doxorubicin in human TNBC xenografts. PEG engagers with specificity for EGFR are promising for improved treatment of EGFR+ TNBC patients.

show abstract

Section: Discussionmentioning

confidence: 99%

Conditional internalization of PEGylated nanomedicines by PEG engagers for triple negative breast cancer therapy

Burnouf

Chuang

et al. 2017

Nat Commun

View full text Add to dashboard Cite

show abstract

“…One should note that the paradigm of sequence motifs dictating interactions between TM helices seems now to be an over-simplification of the complex nature of interactions in the membrane (57). Mutating residues in the TMD of EGFR did affect clustering and basal and EGF-induced endocytosis but not EGF-induced kinase activity (58). Interestingly, an NMR structure of the TMD and flanking regions (59) suggests the helixhelix packing to be mediated by the N-terminal TM helix sequence motif, which is not conserved in the TMD of EGFR.…”

Section: Discussionmentioning

confidence: 99%

Type I Interferon Signaling Is Decoupled from Specific Receptor Orientation through Lenient Requirements of the Transmembrane Domain

Sharma

Longjam

Schreiber

2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Type I interferons serve as the first line of defense against pathogen invasion. Binding of IFNs to its receptors, IFNAR1 and IFNAR2, is leading to activation of the IFN response. To determine whether structural perturbations observed during binding are propagated to the cytoplasmic domain, multiple mutations were introduced into the transmembrane helix and its surroundings. Insertion of one to five alanine residues near either the N or C terminus of the transmembrane domain (TMD) likely promotes a rotation of 100°and a translation of 1.5 Å per added residue. Surprisingly, the added alanines had little effect on the binding affinity of IFN to the cell surface receptors, STAT phosphorylation, or gene induction. Similarly, substitution of the juxtamembrane residues of the TMD with alanines, or replacement of the TMD of IFNAR1 with that of IFNAR2, did not affect IFN binding or activity. Finally, only the addition of 10 serine residues (but not 2 or 4) between the extracellular domain of IFNAR1 and the TMD had some effect on signaling. Bioinformatic analysis shows a correlation between high sequence conservation of TMDs of cytokine receptors and the ability to transmit structural signals. Sequence conservation near the TMD of IFNAR1 is low, suggesting limited functional importance for this region. Our results suggest that IFN binding to the extracellular domains of IFNAR1 and IFNAR2 promotes proximity between the intracellular domains and that differential signaling is a function of duration of activation and affinity of binding rather than specific conformational changes transmitted from the outside to the inside of the cell.Type I interferons (IFNs) orchestrate the antiviral innate immunity in vertebrates. They consist of 16 members in humans as follows: 12 different IFN␣ subtypes, as well as IFN␤, IFN, IFN⑀, and IFN. They are clustered on the short arm of chromosome 9. All type I interferons elicit their innate and adaptive immune responses after binding to the receptor and forming the IFNAR1-IFN-IFNAR2 ternary complex (1, 2), the formation of which is essential for signaling. Both IFNAR1 and IFNAR2 belong to the class II helical cytokine receptors with four fibronectin type III-like subdomains for IFNAR1 and two subdomains for IFNAR2. The extracellular domains are followed by a single helix of 21 amino acids spanning the membrane, which in turn is connected to mostly natively unstructured intracellular domains and their associated effectors (Fig. 1A) (3). Cells lacking one of the receptors lack normal IFN signaling (4, 5). Structurally different members of type I IFNs bind to the same cell surface receptor but mediate differential responses that result from differences in binding affinities, concentration of IFN, and duration of activation (5-13). Differential signaling is realized through robust (antiviral) versus tunable (antiproliferative and immunomodulatory) gene induction leading to different phenotypic outcomes (14,15). Receptor dimerization drives the activation of cytosolic associated Janus family kina...

show abstract

“…Although nanobodies distribute and bind to their targets very rapidly, the background signal is relatively high during the first hours. Monovalent nanobodies are not efficiently internalized and therefore bound nanobody can unbind over a certain period of time . The 7D12–800CW washout from normal tissue occurs faster than the release from its receptor in the tumor.…”

Section: Discussionmentioning

confidence: 99%

Intraoperative fluorescence delineation of head and neck cancer with a fluorescent Anti‐epidermal growth factor receptor nanobody

Driel

Vorst

Verbeek

et al. 2013

Intl Journal of Cancer

View full text Add to dashboard Cite

Intraoperative near-infrared (NIR) fluorescence imaging is a technology with high potential to provide the surgeon with real-time visualization of tumors during surgery. This study explores the feasibility for clinical translation of an epidermal growth factor receptor (EGFR) targeting nanobody for intraoperative imaging and resection of orthotopic tongue tumors and cervical lymph node metastases. The anti-EGFR nanobody 7D12 and the negative control nanobody R2 were conjugated to the NIR fluorophore IRDye800CW (7D12-800CW and R2-800CW). Orthotopic tongue tumors were induced in nude mice using the OSC-19-luc2-cGFP cell line. Tumor bearing mice were injected with 25µg 7D12-800CW, R2-800CW or 11µg 800CW. Subsequently, other mice were injected with 50 µg or 75 µg of 7D12-800CW. The FLARE imaging system and the IVIS spectrum were used to identify, delineate and resect the primary tumor and cervical lymph node metastases. All tumors could be clearly identified using 7D12-800CW. A significantly higher tumor-to-background ratio (TBR) was observed in mice injected with 7D12-800CW compared to mice injected with R2-800CW and 800CW. The highest average TBR (2.00 ± 0.34 and 2.72 ± 0.17 for FLARE and IVIS spectrum, respectively) was observed 24 hours after administration of the EGFR-specific nanobody. After injection of 75 µg 7D12-800CW cervical lymph node metastases could be clearly detected. Orthotopic tongue tumors and cervical lymph node metastases in a mouse model were clearly identified intraoperatively using a recently developed fluorescent EGFR targeting nanobody. Translation of this approach to the clinic would potentially improve the rate of radical surgical resections.

show abstract

EGFR endocytosis requires its kinase activity and N-terminal transmembrane dimerization motif

Cited by 54 publications

References 75 publications

Conditional internalization of PEGylated nanomedicines by PEG engagers for triple negative breast cancer therapy

Conditional internalization of PEGylated nanomedicines by PEG engagers for triple negative breast cancer therapy

Type I Interferon Signaling Is Decoupled from Specific Receptor Orientation through Lenient Requirements of the Transmembrane Domain

Intraoperative fluorescence delineation of head and neck cancer with a fluorescent Anti‐epidermal growth factor receptor nanobody

Contact Info

Product

Resources

About