Protein aliquots were separated on SDS-PAGE gels, transferred to PVDF membranes, blocked with 3% non-fat milk, and then incubated overnight with rabbit polyclonal antibodies against FGFR1 (sc-121, 1:400) and FGFR2
(sc-122, 1:600) (Santa Cruz Biotechnology, Santa Cruz, CA), respectively.
showed that the inhibition of FGFR1 and FGFR2
increased cisplatin sensitivity in ovarian cancer (36).
23] demonstrated that the variants in FGFR2
associated with breast cancer risk were associated with increased expression of FGFR2
, secondary to altered binding of transcription factors Oct-1/Runx2 and C/EBP?
Most of the remaining significantly mutated genes (PTEN, PIK3CA, PIK3R1, ARID1A, RPL22, KRAS, CTNNB1, ATR, FGFR2
, CCND1) have well-documented roles in the endometrioid subtype, as discussed earlier in this review and elsewhere (24, 65).
Consistent with this effort at AVEO, recent data from human correlative and mutational studies has implicated FGFR2
as a potentially important target in human breast cancer.
In particular, mutations in EGFR, BRAF, MET, and FGFR2
may have important therapeutic implications for patients with these tumor subtypes.
8] Human genes: KLK3, kallikrein-related peptidase 3; TERT, telomerase reverse transcriptase; FGFR2
, fibroblast growth factor receptor 2; TBBX3, T-box protein 3; KLK15, kallikrein-related peptidase 15; KLK2, kallikrein-related peptidase 2; KLK1, kallikrein-related peptidase 1; ACPT, acid phosphatase, testicular; GPR32, G protein-coupled receptor 32; INS, insulin; KLK4, kallikrein-related peptidase 4; KLK9, kallikrein-related peptidase 9; KLK10, kallikrein-related peptidase 10; KLK14, kallikrein-related peptidase 14.
Highlights from these presentations include preclinical data regarding AV-951, a novel, triple-VEGFR inhibitor, as a potential treatment option for hepatocellular carcinoma (HCC); preclinical activity of SCH 900105 (AV-299), the first candidate from AVEO's robust antibody pipeline to enter clinical development targeting the c-Met pathway; and the identification of FGFR2
as a compelling new target in the treatment of various human cancers, via AVEO's propriety genetic screens.
Other SNPs that reached genome-wide significance were rs401681 and rs2736098 near the TERT (telomerase reverse transcriptase) gene on chromosome 5, rs10788160 near the FGFR2
(fibroblast growth factor receptor 2) gene, rs10993994 near the MSMB (microseminoprotein, beta-) gene (as in prior studies) on chromosome 10, rs11067228 near the TBX3 (T-box 3) gene on chromosome 12, and rs4430796 at the HNF1B (HNF1 homeobox B) gene on chromosome 17.
Specific cancer genes overexpressed among tumors in the study's cohort included: ALK, AR, ARAF, BRAF, FGFR2
, GLI1, GLI2, HRAS, HSP90AA1, KRAS, MET, NOTCH2, NOTCH3, and SHH.
Most of the mutations have been found in FGFR2
Multiplex quantitative PCR was used to amplify the FGFR2
, KRIT1, and SNRPN genes (FGFR2
forward, CAC AAT CAT TCC TGT GTC GT; FGFR2
reverse, AGC AGT CAA CCA AGA AAA GG; KRIT1 forward, TTC GAA TGG CTA CTT CTA CCT G; KRIT1 reverse, AAA ACG TCT TTT AAA TCA GAG C; SNRPN forward, CTT TGT ACT CCT CCA GCA AC; SNRPN reverse, TAC AGG AAT GAA AGG CAT TA).