An activated form of ADAM10 is tumor selective and regulates cancer stem-like cells and tumor growth

Atapattu, Lakmali; Saha, Nayanendu; Chheang, Chanly; Eissman, Moritz F.; Xu, Kai; Vail, Mary E.; Hii, Linda; Llerena, Carmen; Liu, Zhanqi; Horvay, Katja; Abud, Helen E.; Kusebauch, Ulrike; Moritz, Robert L.; Ding, Bi-Sen; Cao, Zhongwei; Rafii, Shahin; Ernst, Matthias; Scott, Andrew M.; Nikolov, Dimitar B.; Lackmann, Martin; Janes, Peter W.

doi:10.1084/jem.20151095

Cited by 56 publications

(74 citation statements)

References 57 publications

(112 reference statements)

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“…The structure likely represents an autoinhibited conformation (see below), because the location of the cysteine-rich domain at the mouth of the peptide-binding groove partially occludes the active site, restricting access of substrate to the specificity pocket for the residue that immediately follows the scissile bond (Figure 1A). The D+C domains contain little secondary structure and adopt a cupped shape, rigidified by a disulfide-bond network and a conserved Ca 2+ -binding loop as in the isolated D+C region of bovine ADAM10 (PDB ID code 5L0Q; (Atapattu et al, 2016)).…”

Section: Resultsmentioning

confidence: 99%

“…To evaluate the autoinhibition model further, we assayed the catalytic activity of the enzyme using a fluorogenic peptide substrate in the presence and absence of the modulatory antibody 8C7. This antibody binds to a site on the cysteine-rich domain, and the x-ray structure (Atapattu et al, 2016) of a complex between the 8C7 F ab and the isolated D+C region of bovine ADAM10, missing the protease domain, has been solved previously (Figure 3C). Superposition of the intact ADAM10 ectodomain onto this structure reveals that the ADAM10 catalytic domain and 8C7 heavy chain would overlap, resulting in a substantial steric clash (Figure 3D).…”

Section: Resultsmentioning

confidence: 99%

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Structural Basis for Regulated Proteolysis by the α-Secretase ADAM10

Seegar

Killingsworth

Saha

et al. 2017

Cell

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SUMMARY Cleavage of membrane-anchored proteins by ADAM (a disintegrin and metalloproteinase) endopeptidases plays a key role in a wide variety of biological signal transduction and protein turnover processes. Among ADAM family members, ADAM10 stands out as particularly important because it is both responsible for regulated proteolysis of Notch receptors and catalyzes the non-amyloidogenic α-secretase cleavage of the Alzheimer’s precursor protein, APP. We present here the X-ray crystal structure of the ADAM10 ectodomain, which together with biochemical and cellular studies reveals how access to the enzyme active site is regulated. The enzyme adopts an unanticipated architecture, in which the C-terminal cysteine-rich domain partially occludes the enzyme active site, preventing unfettered substrate access. Binding of a modulatory antibody to the cysteine-rich domain liberates the catalytic domain from autoinhibition, enhancing enzymatic activity toward a peptide substrate. Together, these studies reveal a mechanism for regulation of ADAM activity and offer a roadmap for its modulation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Structural Basis for Regulated Proteolysis by the α-Secretase ADAM10

Seegar

Killingsworth

Saha

et al. 2017

Cell

Self Cite

124

135

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show abstract

“…It is possible that the previously described antimetastasis mechanism of action of aurora kinase inhibitors is predominantly driving this phenotype (41). Although the combination of in vitro RNAi studies in 3D spheroids suggests cooperativity between the individual targets of inhibitor #1, their individual impact might be different in vivo (47,48).…”

Section: Discussionmentioning

confidence: 99%

Target Deconvolution of a Multikinase Inhibitor with Antimetastatic Properties Identifies TAOK3 as a Key Contributor to a Cancer Stem Cell–Like Phenotype

Bian

Teper

Griner

et al. 2019

Molecular Cancer Therapeutics

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Pancreatic cancer remains an incurable condition. Its progression is driven, in part, by subsets of cancer cells that evade the cytotoxic effects of conventional chemotherapies. These cells are often low-cycling, multidrug resistant, and adopt a stem cell-like phenotype consistent with the concept of cancer stem cells (CSC). To identify drugs impacting on tumor-promoting CSCs, we performed a differential high-throughput drug screen in pancreatic cancer cells cultured in traditional (2D) monolayers versus threedimensional (3D) spheroids which replicate key elements of the CSC model. Among the agents capable of killing cells cultured in both formats was a 1H-benzo[d]imidazol-2amine-based inhibitor of IL2-inducible T-cell kinase (ITK; NCGC00188382, inhibitor #1) that effectively mediated growth inhibition and induction of apoptosis in vitro, and suppressed cancer progression and metastasis formation in vivo. An examination of this agent's polypharmacology via in vitro and in situ phosphoproteomic profiling demonstrated an activity profile enriched for mediators involved in DNA damage repair. Included was a strong inhibitory potential versus the thousand-and-one amino acid kinase 3 (TAOK3), CDK7, and aurora B kinases. We found that cells grown under CSC-enriching spheroid conditions are selectively dependent on TAOK3 signaling. Loss of TAOK3 decreases colony formation, expression of stem cell markers, and sensitizes spheroids to the genotoxic effect of gemcitabine, whereas overexpression of TAOK3 increases stem cell traits including tumor initiation and metastasis formation. By inactivating multiple components of the cell-cycle machinery in concert with the downregulation of key CSC signatures, inhibitor #1 defines a distinctive strategy for targeting pancreatic cancer cell populations.

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“…This, paired with improved knowledge of tetraspanin regulation of ADAM10, may allow generation of better inhibitors that are able to specifically target ADAM10's cleavage of particular substrates (6). Further, protease inhibitors may be surpassed in specificity through the use of monoclonal antibodies that mask the binding pocket for ADAM10, like mAb 8C7 (135). Yet, without the implementation of a personalized medicine approach, especially in cancer, ADAM10 inhibition therapy will most likely continue to fail in clinical trials.…”

Section: Discussionmentioning

confidence: 99%