Inhibitor-induced HER2-HER3 heterodimerisation promotes proliferation through a novel dimer interface

Claus, Jeroen; Patel, Gargi; Autore, Flavia; Colomba, Audrey; Weitsman, Gregory; Soliman, Tanya; Roberts, Selene K.; Zanetti-Domingues, Laura C.; Hirsch, Michael; Collu, Francesca; George, Roger; Ortiz-Zapater, Elena; Barber, Paul R.; Vojnovic, B.; Yarden, Yosef; Martin-Fernandez, Marisa L.; Cameron, Angus J.M.; Fraternali, Franca; Ng, Tony; Parker, Peter J.

doi:10.7554/elife.32271

Cited by 57 publications

(57 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, the total levels of Her2, Her3 and EGFR in Her2 nanoenvironments were higher in SKBR3 cells compared to MCF7 cells. These results are consistent with previous reports showing that heterodimers between Her2 and other EGFR family members can form in Her2-expressing cancer cells, irrespective of Her2 expression levels 16,19 . Together, these results support the notion that higher expression levels of Her2 lead to a higher number of interactions between Her2 and other EGF receptors resulting in increased oncogenic signalling in Her2overexpressing cells compared to cancer cells with basal expression levels of Her2.…”

Section: Her2supporting

confidence: 93%

“…Interestingly, the oncogenic capacity of Her2 is closely connected to the impact of overexpression on the frequency distributions of interactions between Her2 and EGFR family members at the cell membrane [14][15][16] . For example, Her2 overexpression leads to increased levels of Her2 and Her3 heterodimers, which drive more potent oncogenic signalling activity than the corresponding homodimers 13,[17][18][19] . Further, Her2-EGFR dimerization, driven by overexpression of one or both proteins, has been shown to lead to a more aggressive breast cancer phenotype 20 .…”

mentioning

confidence: 99%

See 1 more Smart Citation

A DNA nanoassembly-based approach to map membrane protein nanoenvironments

Ambrosetti

Bernardinelli

Hoffecker

et al. 2019

Preprint

View full text Add to dashboard Cite

Super-resolution imaging has revealed that most proteins at the plasma membrane are not uniformly distributed but localize to dynamic domains of nanoscale dimensions. To investigate their functional relevance, there is a need for methods that enable comprehensive mapping of the compositions and spatial organizations of membrane protein nanodomains in cell populations. However, current superresolution methods are limited to analysing small, preselected subsets of proteins, at very low sampling fractions. Here we describe the development of a non-microscopy based super-resolution method for unbiased ensemble analysis of membrane protein nanodomains. The method, termed NANOscale DEciphEring of membrane Protein nanodomains (NanoDeep), is based on the use of DNA nanoassemblies to translate membrane protein organization information into a DNA sequencing readout. Using NanoDeep, we characterized the nanoenvironments of Her2, a membrane receptor of critical relevance in cancer. We found that the occupancies of Her2, Her3 and EGFR in the nanoenvironments surrounding Her2 were similar in two cell lines with vastly different expression levels of Her2. Further, we found that adding Heregulin-β1 to cancer cells led to increased occupancy of Her2 and Her3, and to a lesser extent EGFR, in Her2 nanoenvironments. NanoDeep has the potential to provide new insights into the roles of the composition and spatial organization of protein nanoenvironments in the regulation of membrane protein function. MainCells sense extracellular signals, such as protein ligands, through specialized proteins present on the cell surface called membrane receptors. The protein nanoenvironment, i.e. the composition and spatial organization of proteins surrounding membrane receptors, is dynamic and often modulated by ligand binding, suggesting that it has functional relevance 1, 2 . Super-resolution microscopy has enabled the characterization of the nanoscale spatial distributions of proteins at the cell membrane but is limited to simultaneously analysing only a few proteins. Further, using super-resolution microscopy, it is only feasible to image a small fraction of all protein nanodomains present in a cell population 3-5 . DNA detection as a proxy for protein detection through the use of oligo-conjugated affinity binders has been used extensively for signal amplification 6 and analysis of proximity between pairs of proteins 7, 8 . Further, DNA sequencing is used as a readout in DNA microscopy, a new method to visualize the spatial organization of RNA and DNA molecules inside cells 9-11 . Here we present NanoDeep, a method that uses DNA sequencing to decipher the nanoscale spatial distribution of membrane proteins. This method allows for the detection en masse of the inventory of proteins that forms the nanoenvironment of any reference membrane protein in cell populations. We demonstrate the application of NanoDeep to the analysis of protein nanoenviroments surrounding Human Epidermal Growth Factor Receptor 2 (Her2). Her2 cooperates with members of the Ep...

show abstract

Section: Her2supporting

confidence: 93%

mentioning

confidence: 99%

A DNA nanoassembly-based approach to map membrane protein nanoenvironments

Ambrosetti

Bernardinelli

Hoffecker

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…6E), which will target 20% or less from the total EGFR pool. Significantly, there are evidences that only for the constitutively untethered HER2, lapatinib stabilizes ligand-responsive dimers (34), associated with tumor progression and resistance (35).…”

Section: Discussionmentioning

confidence: 99%

Oncogenic mutations at the EGFR ectodomain structurally converge to remove a steric hindrance on a kinase-coupled cryptic epitope

Orellana

Thorne

Lema

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Epidermal growth factor receptor (EGFR) signaling is initiated by a large ligand-favored conformational change of the extracellular domain (ECD) from a closed, self-inhibited tethered monomer, to an open untethered state, which exposes a loop required for strong dimerization and activation. In glioblastomas (GBMs), structurally heterogeneous missense and deletion mutations concentrate at the ECD for unclear reasons. We explore the conformational impact of GBM missense mutations, combining elastic network models (ENMs) with multiple molecular dynamics (MD) trajectories. Our simulations reveal that the main missense class, located at the I-II interface away from the self-inhibitory tether, can unexpectedly favor spontaneous untethering to a compact intermediate state, here validated by smallangle X-ray scattering (SAXS). Significantly, such intermediate is characterized by the rotation of a large ECD fragment (N-TR1), deleted in the most common GBM mutation, EGFRvIII, and that makes accessible a cryptic epitope characteristic of cancer cells. This observation suggested potential structural equivalence of missense and deletion ECD changes in GBMs. Corroborating this hypothesis, our FACS, in vitro, and in vivo data demonstrate that entirely different ECD variants all converge to remove N-TR1 steric hindrance from the 806epitope, which we show is allosterically coupled to an intermediate kinase and hallmarks increased oncogenicity. Finally, the detected extraintracellular coupling allows for synergistic cotargeting of the intermediate with mAb806 and inhibitors, which is proved herein. cancer | mutational heterogeneity | structural convergence | intermediate | cryptoepitope

show abstract

“…In fact, one of the determinants of HER3's lack of activity is the absence of an intact receiver interface. A recent study proposed that under certain circumstances, such as the presence of lapatinib that binds HER2 with high affinity, the HER2/HER3 KD heterodimer may also adopt a head‐to‐head orientation upon stimulation with the HER3 ligand, neuregulin (NRG) . This interface was first observed in the crystal lattice of the HER3 KD structure supporting head‐to‐head HER3 homodimers .…”

Section: Egfr: Breaking Symmetry For Activationmentioning

confidence: 99%

More than the sum of the parts: Toward full‐length receptor tyrosine kinase structures

2019

View full text Add to dashboard Cite

Intercellular communication governs complex physiological processes ranging from growth and development to the maintenance of cellular and organ homeostasis. In nearly all metazoans, receptor tyrosine kinases (RTKs) are central players in these diverse and fundamental signaling processes. Aberrant RTK signaling is at the root of many developmental diseases and cancers and it remains a key focus of targeted therapies, several of which have achieved considerable success in patients. These therapeutic advances in targeting RTKs have been propelled by numerous genetic, biochemical, and structural studies detailing the functions and molecular mechanisms of regulation and activation of RTKs. The latter in particular have proven to be instrumental for the development of new drugs, selective targeting of mutant forms of RTKs found in disease, and counteracting ensuing drug resistance. However, to this day, such studies have not yet yielded high‐resolution structures of intact RTKs that encompass the extracellular and intracellular domains and the connecting membrane‐spanning transmembrane domain. Technically challenging to obtain, these structures are instrumental to complete our understanding of the mechanisms by which RTKs are activated by extracellular ligands and of the effect of pathological mutations that do not directly reside in the catalytic sites of tyrosine kinase domains. In this review, we focus on the recent progress toward obtaining such structures and the insights already gained by structural studies of the subdomains of the receptors that belong to the epidermal growth factor receptor, insulin receptor, and platelet‐derived growth factor receptor RTK families. © 2019 IUBMB Life, 71(6):706–720, 2019

show abstract

Inhibitor-induced HER2-HER3 heterodimerisation promotes proliferation through a novel dimer interface

Cited by 57 publications

References 87 publications

A DNA nanoassembly-based approach to map membrane protein nanoenvironments

A DNA nanoassembly-based approach to map membrane protein nanoenvironments

Oncogenic mutations at the EGFR ectodomain structurally converge to remove a steric hindrance on a kinase-coupled cryptic epitope

More than the sum of the parts: Toward full‐length receptor tyrosine kinase structures

Contact Info

Product

Resources

About