RET is constitutively activated by novel tandem mutations that alter the active site resulting in multiple endocrine neoplasia type 2B

Aaron N. Cranston, Cristiana Carniti, Kim Oakhill, Elzbieta Radzio-Andzelm, Eric A. Stone, Andrew S McCallion, Shirley Hodgson, Sue Clarke, Piera Mondellini, Jean Leyland, Marco A. Pierotti, Joanne Whittaker, Susan S. Taylor, Italia Bongarzone, Bruce A J Ponder

Research output: Contribution to journalArticle

Abstract

Constitutive activation of the RET receptor tyrosine kinase underlies the genesis and progression of multiple endocrine neoplasia type 2 (MEN 2), a dominantly inherited cancer predisposition. Importantly, although kinase activation represents a common theme in neoplasias, not all activating mutations are functionally equivalent. Consistent with this, we ascertained a patient with classical features of MEN 2B, but lacking either of the classical mutations in RET (M918T or A883F). Instead, the patient harbors a novel pair of germ line missense mutations in cis at codons 804 and 805. We evaluated the potential physiochemical effects of these substitutions in silico, predicting both to be moderately deleterious in isolation, but severely deleterious in combination. Consistent with this postulate, we show that the identified tandem mutations (V804M/E805K) are biologically active, transforming cells in culture and that their transforming capacity in combination is distinctly synergistic. Furthermore, the V804M/E805K tandem lesion confers resistance to the small molecule receptor tyrosine kinase inhibitor, PP1, suggesting a mode of action distinct from that known for classical MEN 2B mutations. To address this question, we used homology molecular modeling in silico to model the active site of RET. We predict that RET804 constitutes a critical gatekeeper residue that, when mutated in combination with RET805, induces a conformational change in the hinge region that locks the active site in a position permissive for ATP hydrolysis. Our findings have implications both in the clinic and in the successful development of novel kinase-targeted anticancer drugs.

Original languageEnglish (US)
Pages (from-to)10179-10187
Number of pages9
JournalCancer Research
Volume66
Issue number20
DOIs
StatePublished - Oct 15 2006

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Multiple Endocrine Neoplasia Type 2b
Catalytic Domain
Mutation
Computer Simulation
Proto-Oncogene Proteins c-ret
Phosphotransferases
Multiple Endocrine Neoplasia Type 2a
Germ-Line Mutation
Missense Mutation
Codon
Neoplasms
Hydrolysis
Cell Culture Techniques
Adenosine Triphosphate
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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RET is constitutively activated by novel tandem mutations that alter the active site resulting in multiple endocrine neoplasia type 2B. / Cranston, Aaron N.; Carniti, Cristiana; Oakhill, Kim; Radzio-Andzelm, Elzbieta; Stone, Eric A.; McCallion, Andrew S; Hodgson, Shirley; Clarke, Sue; Mondellini, Piera; Leyland, Jean; Pierotti, Marco A.; Whittaker, Joanne; Taylor, Susan S.; Bongarzone, Italia; Ponder, Bruce A J.

In: Cancer Research, Vol. 66, No. 20, 15.10.2006, p. 10179-10187.

Research output: Contribution to journalArticle

Cranston, AN, Carniti, C, Oakhill, K, Radzio-Andzelm, E, Stone, EA, McCallion, AS, Hodgson, S, Clarke, S, Mondellini, P, Leyland, J, Pierotti, MA, Whittaker, J, Taylor, SS, Bongarzone, I & Ponder, BAJ 2006, 'RET is constitutively activated by novel tandem mutations that alter the active site resulting in multiple endocrine neoplasia type 2B', Cancer Research, vol. 66, no. 20, pp. 10179-10187. https://doi.org/10.1158/0008-5472.CAN-06-0884
Cranston, Aaron N. ; Carniti, Cristiana ; Oakhill, Kim ; Radzio-Andzelm, Elzbieta ; Stone, Eric A. ; McCallion, Andrew S ; Hodgson, Shirley ; Clarke, Sue ; Mondellini, Piera ; Leyland, Jean ; Pierotti, Marco A. ; Whittaker, Joanne ; Taylor, Susan S. ; Bongarzone, Italia ; Ponder, Bruce A J. / RET is constitutively activated by novel tandem mutations that alter the active site resulting in multiple endocrine neoplasia type 2B. In: Cancer Research. 2006 ; Vol. 66, No. 20. pp. 10179-10187.
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AU - Cranston, Aaron N.

AU - Carniti, Cristiana

AU - Oakhill, Kim

AU - Radzio-Andzelm, Elzbieta

AU - Stone, Eric A.

AU - McCallion, Andrew S

AU - Hodgson, Shirley

AU - Clarke, Sue

AU - Mondellini, Piera

AU - Leyland, Jean

AU - Pierotti, Marco A.

AU - Whittaker, Joanne

AU - Taylor, Susan S.

AU - Bongarzone, Italia

AU - Ponder, Bruce A J

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