Fibroblast Growth Factor Receptor 4 Gene Research Articles

Role of serum endotoxin, FGF19, TLR2, TNF-α, IL-12 and IL-10 in NAFLD-associated T2DM pathogenesis: Insights into Th1 bias and protective mechanisms.

Non-alcoholic fatty liver disease (NAFLD) in non-obese patients is pathophysiologically distinct, exhibiting common immunological link with type-2 diabetes mellitus (T2DM). This study aims to delineate the role of Toll-like receptor 2 (TLR2)-mediated immuno-modulation along with its association with fibroblast growth factor receptor 4 (FGFR4) and its ligand fibroblast growth factor 19 (FGF19) in the pathogenesis of NAFLD without or with T2DM. Blood samples were collected from patients with NAFLD (n = 90), NAFLD with T2DM (n = 90) and healthy cohorts (n = 90) with consent and clinical records. Real-time polymerase chain reaction (PCR), enzyme-linked immunoassay (ELIZA) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to analyze messenger ribonucleic acid (mRNA), protein expression and gene polymorphism. The molecular genetic analysis revealed the prevalence of variant allele(A) in FGFR4 gene in both cases compared to controls. The mRNA expression of FGF19 and TLR2 exhibited significant upregulation in NAFLD without T2DM compared to NAFLD with T2DM. Tumor necrosis factor-α (TNF-α) and interleukin-12 (IL-12) showed upregulation in both disease cohorts compared to control while IL-10 showed significant downregulation in NAFLD with T2DM compared to the other two cohorts. Correlation analysis between FGF19 and TLR2 revealed significant positive association in both NAFLD with and without T2DM. The Th1:Th2 ratio showed significant upregulation in NAFLD with T2DM compared to NAFLD without T2DM. In conclusion, elevated serum endotoxin levels appear to contribute to NAFLD and T2DM development. Upregulated FGF19 seems to be protective against developing T2DM in NAFLD patients. Higher TLR2, TNF-α and IL-12 expression in NAFLD without T2DM suggests a Th1 bias in its pathogenesis, while reduced IL-10 in NAFLD with T2DM implies a more skewed Th1 state in this condition.

Influence of temperature on embryonic development of Pontastacus leptodactylus freshwater crayfish, and characterization of growth and osmoregulation related genes

Narrow clawed crayfish, Pontastacus (Astacus) leptodactylus, represents an ecologically and economically valuable freshwater species. Despite the high importance of artificial breeding for conservation purpose and aquaculture potential, hatching protocols have not been developed so far in this species. Further, limited knowledge exists regarding the artificial egg incubation, the temperature effect on embryonic development, hatching synchronization and hatching rate. In the present study we investigated the temperature increase (from 17 oC to 22oC) effects in two different embryonic developmental stages of P. leptodactylus. Furthermore, two primer pairs for the Fibroblast Growth Factor Receptor 4 (FGFR4) gene cDNA amplification were successfully designed, characterising for the first time the FGFR4 gene in P. leptodactylus in relation to different developmental stages and temperatures. Apart from the FGFR4 gene, the Na+/K+-ATPase α-subunit expression was also explored. Both the FGFR4 and Na+/K+-ATPase α-subunit expression levels were higher in embryos closer to hatching. Egg incubation at 22oC for seven days led to significant increase of FGFR4 expression in embryos from earlier developmental stages. Nevertheless, temperature increase did not affect FGFR4 expression in eggs from latter developmental stages and Na+/K+-ATPase α-subunit expression in all developmental stages. Temperature increase represents therefore probably a promising strategy for accelerating hatching in freshwater crayfish particularly in early developmental stages. Specifically, our results indicate that FGFR4 expression increased in embryonic stages closer to hatching and that temperature influences significantly its expression in embryos from earlier developmental stages. Overall, these findings can provide a better understanding of artificial egg incubation of P. leptodactylus, and therefore can be employed for the effective management of this species, both for economic and biodiversity retention reasons.

Impact of FGFR4 Gene Polymorphism on the Progression of Colorectal Cancer.

Colorectal cancer (CRC) is a multifactorial malignancy, and its high incidence and mortality rate remain a global public health burden. Fibroblast growth factor receptor 4 (FGFR4) is a receptor tyrosine kinase that has been shown to play a key role in cancer development and prognosis via the activation of its downstream oncogenic signaling pathways. The present study aimed to explore the impact of FGFR4 gene polymorphisms on the risk and progression of CRC. Three FGFR4 single-nucleotide polymorphisms (SNPs), including rs1966265, rs351855, and rs7708357, were evaluated in 413 CRC cases and 413 gender- and age-matched cancer-free controls. We did not observe any significant association of three individual SNPs with the risk of CRC between the case and control group. However, while assessing the clinicopathological parameters, patients of rectal cancer possessing at least one minor allele of rs1966265 (AG and GG; AOR, 0.236; p = 0.046) or rs351855 (GA and AA; AOR, 0.191; p = 0.022) were found to develop less metastasis as compared to those who are homozygous for the major allele. Further analyses using the datasets from the Genotype-Tissue Expression (GTEx) Portal and The Cancer Genome Atlas (TCGA) revealed that rs351855 regulated FGFR4 expression in many human tissues, and increased FGFR4 levels were associated with the occurrence, advanced stage, and distal metastasis of colon adenocarcinoma. These data suggest that the amino acid change in combination with altered expression levels of FGFR4 due to genetic polymorphisms may affect CRC progression.

FGFR4 Gene Polymorphism Reduces the Risk of Distant Metastasis in Lung Adenocarcinoma in Taiwan.

Fibroblast growth factor receptor 4 (FGFR4) is involved in multiple physiological and pathological processes. Several genetic variants of FGFR4 have been shown to be associated with tumor progression in many cancers. However, its association, such as genetic variants and expression levels, with lung cancer is controversial. The present study examined the relationship between four single-nucleotide polymorphisms (SNPs; rs2011077 T/C, rs351855 G/A, rs7708357 G/A, and rs1966265 A/G) of FGFR4 and the risk of lung adenocarcinoma with the epidermal growth factor receptor (EGFR) mutation status in a Taiwanese cohort. The results demonstrated that FGFR4 rs2011077 (odds ratio (OR) = 0.348, 95% confidence interval (CI) = 0.136–0.891, p = 0.024), and rs351855 (OR = 0.296, 95% CI = 0.116–0.751, p = 0.008) showed an inverse association with distant metastasis in wild-type EGFR lung adenocarcinoma. Furthermore, a database analysis using The Cancer Genome Atlas revealed that the higher FGFR4 expression level was correlated with poor survival rates in wild-type EGFR lung adenocarcinoma. In conclusion, the data suggest that FGFR4 SNPs may help in identifying patient subgroups at low-risk for tumor metastasis, among carriers of lung adenocarcinoma bearing wild-type EGFR.

Involvement of FGFR4 Gene Variants on the Clinicopathological Severity in Urothelial Cell Carcinoma.

Fibroblast growth factor receptor 4 (FGFR4) plays a prominent role in cell proliferation and cancer progression. This study explored the effect of FGFR4 single-nucleotide polymorphisms (SNPs) on the clinicopathological characteristics of urothelial cell carcinoma (UCC). This study was conducted to survey the possible correlation of the polymorphism of FGFR4 to the risk and clinicopathologic characteristics of UCC. Four loci of FGFR4 (rs2011077 T > C, rs351855 G > A, rs7708357 G>A, and rs1966265 A > G) were genotyped via the TaqMan allelic discrimination approach in 428 UCC cases and 856 controls. The results indicated that UCC subjects who carried the SNP rs2011077 TC+CC genotypes were significantly related to a higher tumor stage (odds ratio (OR): 1.751, 95% confidence interval (CI): 1.078–2.846), primary tumor size (OR: 1.637, 95% CI: 1.006–2.662), and histopathologic grading (OR: 1.919, 95% CI: 1.049–3.511). Moreover, the SNP rs1966265 AG+GG genotypes were prominently related to a higher tumor stage (OR: 1.769, 95% CI: 1.082–2.891), primary tumor size (OR: 1.654, 95% CI: 1.011–2.706), and histopathologic grading (OR: 2.006, 95% CI: 1.096–3.674) compared to individuals with AA homozygotes. In conclusion, our data reveal association of FGFR4 polymorphisms with UCC clinicopathologic characteristics. FGFR4 polymorphisms may serve as a marker or therapeutic target in UCC development.

Prognostic Role of the FGFR4-388Arg Variant in Lung Squamous-Cell Carcinoma Patients With Lymph Node Involvement

BackgroundThe identification of prognostic biomarkers for lung squamous-cell carcinoma (SCC) pathology is crucial because of its poor prognosis. A variant of the FGFR4 (fibroblast growth factor receptor 4) gene, FGFR4-388Arg, has been associated with prognosis and is linked to oncogenesis in vitro in several types of cancer. We analyzed the association of this variant with prognosis and downstream signaling alteration in lung SCC patients. MethodsThe presence of the FGFR4-388Arg variant was determined in 114 formalin-fixed, paraffin-embedded lung SCC tissue samples by DNA genotyping and was correlated with clinicopathologic data. The activation of the protein kinase B (AKT) and mitogen-activated protein kinase (MAPK) pathways was determined by immunohistochemistry, and its association with the presence of FGFR4-388Arg was analyzed. ResultsWe found that tumor differentiation status and adjuvant chemotherapy administration could be independent prognostic factors for overall survival (OS) in lymph node–affected patients, as expected. The progression-free survival and OS of patients with lymph node involvement (n = 41) and the FGFR4-388Arg genotype were significantly lower than those of patients lacking this variant (P = .035 and P = .042, respectively). Importantly, multivariate analysis supported the independent prognostic role of the FGFR4-388Arg genotype in OS (P = .025). Regarding downstream signaling, the FGFR4-388Arg genotype was not correlated with altered AKT signaling but was associated with increased MAPK activation in the SCC tumor samples (P = .017). ConclusionThe FGFR4-388Arg variant may represent a promising prognostic biomarker in SCC patients with lymph node involvement. For these patients, FGFR4 may be a potential therapeutic target.

Functional FGFR4 Gly388Arg polymorphism contributes to cancer susceptibility: Evidence from meta-analysis.

Fibroblast growth factor receptor 4 (FGFR4) is a member of receptor tyrosine kinase family. A functional Gly388Arg (rs351855 G>A) polymorphism in FGFR4 gene causes a glycine-to-arginine change at codon 388 within the transmembrane domain of the receptor. Although the FGFR4 rs351855 G>A polymorphism has been implicated in cancer development, its association with cancer risk remains controversial. Here, we have systematically analyzed the association between the rs351855 G>A polymorphism and cancer risk by performing a meta-analysis of 27 studies consisting of 8,682 cases and 9,731 controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to measure the strength of the association. The rs351855 G>A polymorphism was associated with an increased cancer risk under the recessive model (OR=1.19, 95% CI=1.01-1.41). Stratified analysis by cancer type indicated the rs351855 G>A polymorphism was associated with an increased risk of breast and prostate cancer, but a decreased risk of lung cancer. This meta-analysis demonstrates the FGFR rs351855 G>A polymorphism is associated with increased cancer risk and suggests it could potentially serve as a chemotherapeutic target or biomarker to screen high-risk individuals.

Abstract 4996: Targeting FGFR4 with monoclonal antibodies as therapeutic agents for the treatment of rhabdomyosarcoma

Abstract Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood with two major subtypes, embryonal (ERMS) and alveolar (ARMS), and current treatment modalities have yielded event free 5-year survival in only 30% of the patients with high-risk disease. Therefore, there is a need for novel strategies to identify and validate clinically relevant targets and therapeutics for the treatment of RMS. The fibroblast growth factor receptor 4 (FGFR4) is a very attractive therapeutic target because: 1) the FGFR4 gene is over expressed in RMS, 2) it is directly transcriptionally induced by the PAX3-FOXO1 fusion oncogene found in ARMS, 3) it is crucial for survival, proliferation, metastasis and drug resistance, 4) activating mutations in the kinase domain lead to aggressive growth and poor survival in patients with ERMS and 5) genetic or pharmacologic inhibition of FGFR4 signaling inhibited tumor growth in vitro and in vivo. Most normal human tissues do not express FGFR4 protein as determined by immunohistochemistry and electrochemiluminesence assay. Based these, we hypothesize that monoclonal antibodies (mAbs) targeting FGFR4 and their derivatives can serve as potential therapeutic agents for the treatment of RMS. We have developed a total of 15 mAbs against human FGFR4 protein from rabbit and mouse (by hybridoma technology), and from human immunoglobulin libraries (by recombinant DNA technology). These mAbs specifically react with human FGFR4 protein in an ELISA and not to the other members of the FGFR family. Flow cytometer analysis demonstrated that they specifically bound to cell surface FGFR4 protein in RMS cell lines although the FGFR4 expression levels was variable in both ARMS and ERMS cell lines. Furthermore, cell surface FGFR4 mediated internalization of the bound antibody upon incubation at 37oC for as little as 30 min and maximum internalization was observed at 2 h. Two of the mouse mAbs tested were able to mediate specific cytotoxicity in FGFR4-expressing cell lines in the presence of an anti-mouse secondary antibody conjugated to cytotoxic drugs (2oADCs), e.g. monomethyl auristatin F (MMAF) or a duocarmycin derivative (DMDM). Dose titration of the anti-FGFR4 mAbs revealed maximum killing at 100 pM indicating their robust activity in vitro, and both ARMS and ERMS cell lines were susceptible to the mAb-mediated cytotoxicity. Ongoing investigations include generation of FGFR4-targeting direct ADCs and evaluation of their efficacy in vivo. Together, these observations support the contention that anti-FGFR4 mAb can be used as a vehicle to deliver a cytotoxic payload in the form of small molecule drugs and toxins. Thus, FGFR-ADCs may serve as potential therapeutic agents for the treatment of patients with RMS, as well other FGFR4-positive cancers including breast, liver, prostate and lung. Citation Format: Sivasubramanian Baskar. Targeting FGFR4 with monoclonal antibodies as therapeutic agents for the treatment of rhabdomyosarcoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4996.

FGFR4 Is a Potential Predictive Biomarker in Oral and Oropharyngeal Squamous Cell Carcinoma

Objective: The aim of this study was to investigate whether fibroblast growth factor receptor 4 (FGFR4) could serve as a potential therapeutic target, prognostic biomarker or biomarker predicting radiotherapy sensitivity in oral squamous cell carcinoma (OSCC) and oropharyngeal squamous cell carcinoma (OPSCC). Methods: FGFR4 immunohistochemistry and FGFR4/CEN5q FISH were performed on tissue microarrays from 212 OSCC and 238 OPSCC patients. FGFR4 genotypes were determined by PCR and DNA sequencing in 76 random OPSCC samples. The response to radiotherapy was evaluated 3 months after the last radiotherapy treatment session by a head and neck radiation oncologist and/or surgeon during clinic visits. The results were correlated to overall survival and response to radiotherapy. Results: The FGFR4 protein was overexpressed in 64% (153/238) of OPSCCs and 41% (87/212) of OSCCs. The FGFR4 gene was amplified in 0.47% (1/212) of OSCCs and 0.42% (1/238) of OPSCCs, and the FGFR4 Gly388Arg polymorphism was detected in 62% (47/76) of OPSCCs. FGFR4 protein expression, FGFR4 gene copy numbers and FGFR4 genotypes were not related to overall survival or response to radiotherapy in OSCC or OPSCC. Conclusion: FGFR4 is frequently overexpressed in OSCC and OPSCC in the absence of gene amplification, and may serve as a potential predictive marker for FGFR4-directed targeted therapy in OSCC and OPSCC.

Investigation of FGFR4 (Gly388Arg) Gene Polymorphism in Primary Lung Cancer Patients

KEYWORDS Lung Cancer. FGFR4 Gene. Polymorphism. Gly388Arg. Fibroblast ABSTRACT Several studies have shown relationships between predisposition to various types of cancer and polymorphisms of the fibroblast growth factor receptor 4 (FGFR4) gene. In the present study, researchers investigated the relationship between primary lung cancer and (PLC) FGFR4 Gly388Arg polymorphism in regard to tendency, histopathologic sub-type, early onset, and metastatic status. The present study included 124 PLC patients and 100 healthy controls. Polymerase chain reaction-restriction fragment length polymorphism (PCR- RFLP) was used to identify gene polymorphism of FGFR4 Gly388Arg. Statistical significance was considered when p 0.05). The findings in this study demonstrated that there was no relationship between polymorphism of FGFR4 Gly388Arg gene and PLC. However, these results should be confirmed in larger studies and in specific histopathological sub-types of PLC.

Investigation of FGFR4 (Gly388Arg) Gene Polymorphism in Primary Lung Cancer Patients

KEYWORDS Lung Cancer. FGFR4 Gene. Polymorphism. Gly388Arg. Fibroblast ABSTRACT Several studies have shown relationships between predisposition to various types of cancer and polymorphisms of the fibroblast growth factor receptor 4 (FGFR4) gene. In the present study, researchers investigated the relationship between primary lung cancer and (PLC) FGFR4 Gly388Arg polymorphism in regard to tendency, histopathologic sub-type, early onset, and metastatic status. The present study included 124 PLC patients and 100 healthy controls. Polymerase chain reaction-restriction fragment length polymorphism (PCR- RFLP) was used to identify gene polymorphism of FGFR4 Gly388Arg. Statistical significance was considered when p 0.05). The findings in this study demonstrated that there was no relationship between polymorphism of FGFR4 Gly388Arg gene and PLC. However, these results should be confirmed in larger studies and in specific histopathological sub-types of PLC.

Abstract 652: Development and characterization of anti-FGFR4 monoclonal antibodies as therapeutic agents for human rhabdomyosarcoma

Abstract Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood with two major subtypes embryonal (ERMS) and alveolar (ARMS), and current treatment modalities have yielded event free 5-year survival in only 30% of the patients with high-risk disease. Therefore, there is an absolute need for novel strategies and to identify and validate clinically relevant targets for the treatment of RMS. The fibroblast growth factor receptor 4 (FGFR4) is a very attractive therapeutic target because: 1) the FGFR4 gene is over expressed in RMS, 2) it is crucial for survival, proliferation, metastasis and drug resistance, 3) activating mutations in the kinase domain lead to aggressive growth and poor survival in patients with alveolar RMS and 4) genetic or pharmacologic inhibition of FGFR4 signaling inhibited tumor growth in vitro and in vivo. Monoclonal antibodies (mAbs) against specific antigens expressed on cancer cell surface have gained importance as potential therapeutic agents that may be used either alone or in combination with chemotherapeutic drugs. We have developed several mAbs against human FGFR4 protein from rabbit and mouse (by hybridoma technology), and from human immunoglobulin libraries (by recombinant DNA technology). In the present study, we report the development and characterization of some of these mAbs. Biacore analysis of these antibodies showed low nM affinity to purified extracellular domain of FGFR4 (FGFR4-ECD). The ability of these mAbs to bind the native molecule was demonstrated by specific binding to RMS cell lines of both subtypes, and dose response curves exhibited higher binding in ARMS cells than ERMS cells. More importantly, significant binding was noticed in freshly isolated tumor cells from a breast metastatic nodule of a patient with ARMS. The anti-FGFR4 mAbs also bound to transfected cell line expressing FGFR4 on the surface, but not the vector control (FGFR4 negative) cell line indicating the specificity of the reaction. Furthermore, cell surface FGFR4 can mediate internalization of the bound antibody upon incubation at 37C for as little as 30 min and maximum internalization was observed at 2 h. Receptor mediated internalization of the bound mAb was inhibited by a chemical inhibitor, and ARMS cells showed more internalization than ERMS. Together, these observations support the contention that anti- FGFR4 mAb can be used as a vehicle to deliver a cytotoxic payload in the form of small molecule drugs and toxins. Ongoing investigations are aimed at developing anti-FGFR4 mAbs and their derivatives as potential therapeutic agents for the treatment of patients with RMS. This study is supported in part by the intramural research program of the National Cancer Institute, National Institutes of Health, grants from St. Baldrick's Foundation and Stand Up To Cancer - St. Baldrick's Pediatric Dream Team Translational Cancer Research Grant, Grant Number SU2C-AACR-DT11-13. Citation Format: Sivasubramanian Baskar, Zhongyu Zhu, Ramon Lorenzo Labitigan, Michelle Ovanesian, Rimas J. Orentas, Samuel Q. Li, Yohe E. Marielle, John Shern, Dimiter S. Dimitrov, John Maris, Crystal Mackall, Khan Javed. Development and characterization of anti-FGFR4 monoclonal antibodies as therapeutic agents for human rhabdomyosarcoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 652. doi:10.1158/1538-7445.AM2014-652

Switch in FGFR3 and -4 Expression Profile During Human Renal Development May Account for Transient Hypercalcemia in Patients With Sotos Syndrome due to 5q35 Microdeletions

Sotos syndrome is a rare genetic disorder with a distinct phenotypic spectrum including overgrowth and learning difficulties. Here we describe a new case of Sotos syndrome with a 5q35 microdeletion, affecting the fibroblast growth factor receptor 4 (FGFR4) gene, presenting with infantile hypercalcemia. We strove to elucidate the evanescent nature of the observed hypercalcemia by studying the ontogenesis of FGFR3 and FGFR4, which are both associated with fibroblast growth factor (FGF) 23-mediated mineral homeostasis, in the developing human kidney. Quantitative RT-PCR and immunohistochemical analyses were used on archival human kidney samples to investigate the expression of the FGFR signaling pathway during renal development. We demonstrated that renal gene and protein expression of both FGFRs increased during fetal development between the gestational ages (GAs) of 14-40 weeks. Yet FGFR4 expression increased more rapidly as compared with FGFR3 (slope 0.047 vs 0.0075, P = .0018). Moreover, gene and protein expression of the essential FGFR coreceptor, Klotho, also increased with a significant positive correlation between FGFR and Klotho mRNA expression during renal development. Interestingly, we found that perinatal FGFR4 expression (GA 38-40 wk) was 7-fold higher as compared with FGFR3 (P = .0035), whereas in adult kidney tissues, FGFR4 gene expression level was more than 2-fold lower compared with FGFR3 (P = .0029), thus identifying a molecular developmental switch of FGFR isoforms. We propose that the heterozygous FGFR4 deletion, as observed in the Sotos syndrome patient, leads to a compromised FGF23 signaling during infancy accounting for transient hypercalcemia. These findings represent a novel and intriguing view on FGF23 mediated calcium homeostasis.

Fibroblast growth factor receptor 4 polymorphism is associated with increased risk and poor prognosis of non-Hodgkin’s lymphoma

Fibroblast growth factor receptor 4 (FGFR4) is expressed in various cell types and plays important roles in regulating immune responses. Evidence has shown that FGFR4 rs351855 (Gly388Arg) polymorphism may act as a risk factor for many diseases. In the current study, we investigated the association between FGFR4 polymorphisms and the susceptibility to non-Hodgkin's lymphoma (NHL) in the Chinese population. Two polymorphisms in the FGFR4 gene (rs351855G/A and rs147603016G/A) were detected by polymerase chain reaction-restriction fragment length polymorphism in 421 NHL cases and 486 healthy controls. Results showed that prevalence of rs351855AA genotype was significantly increased in patients than in controls (odds ratio [OR] = 2.02, 95% confidence interval [CI] 1.91-3.23, P < 0.001). Similarly, rs351855A allele presented significantly higher numbers in cases compared to healthy donors (49.8 versus 40.1%, P < 0.001). Further study revealed that the frequency of the rs351855G/A polymorphism was clearly elevated in cases with B cell subtype than those with T cell subtypes. When analyzing the survival time of NHL patients with FGFR4 rs351855G/A polymorphism, cases with AA genotype had significantly shorter survival time compared to the patients with GG genotype (P < 0.001) or GA genotype (P < 0.001). These results suggest that FGFR4 rs351855G/A polymorphism is associated with increased susceptibility to NHL and could be used as a marker for predicting the prognosis of the malignancy.

FGFR4 genetic polymorphisms determine the chemotherapy response of Chinese patients with non-small cell lung cancer

To investigate the relationship of fibroblast growth factor receptor 4 (FGFR4) gene polymorphisms with the response of Chinese patients with non-small cell lung cancer (NSCLC) to chemotherapy. A total of 629 patients with Stage III (A+B) or IV NSCLC, as well as 729 age- and gender-matched healthy controls were recruited. All the patients received platinum-based chemotherapy, and the therapeutic effects were evaluated. Three polymorphisms in the FGFR4 gene (rs351855G/A, rs145302848C/G, and rs147603016G/A) were genotyped, and the association between the 3 polymorphisms and the chemotherapy effect was analyzed using SPSS software, version 16.0. The genotype frequencies of rs145302848C/G and rs147603016G/A were not significantly different between NSCLC patients and healthy controls on one hand, and between the responders and non-responders to the chemotherapy on the other hand. The distribution of AA genotype and A-allele of rs351855G/A was significantly lower in NSCLC patients than in healthy controls. Using patients with the GG genotype as a reference, the AA carrier had a significantly reduced risk for the development of NSCLC after normalizing to age, sex and smoking habits. In NSCLC patients, this genotype occurred more frequently in the responders to the chemotherapy than in non-responders. The chance of being a responder was significantly increased with the AA genotype as compared to G genotype. The AA genotype of rs351855G/A had a better prognosis compared with GA and GG genotype carriers: the overall survival of patients with the AA genotype of rs351855G/A was significantly longer than those with the GG+GA genotype (21.1 vs 16.5 months). The rs351855G/A polymorphisms of FGFR4 gene can be used to predict the occurrence, chemotherapy response and prognosis of NSCLC.

Fibroblast growth factor receptor 4 Gly388Arg polymorphism associated with severity of gallstone disease in a Chinese population

The etiology of gallstone disease is multifactorial; supersaturation of bile with cholesterol is a primary cause for gallstone formation. In previous studies, we found that fibroblast growth factor receptor 4 (FGFR4) plays an important role in maintaining bile acid homeostasis by regulating the expression of cholesterol 7α-hydroxylase (CYP7A1), a rate-limiting enzyme for bile acid biosynthesis. The Gly388Arg (G-388R) polymorphism of FGFR4 affects stabilization and activation of FGFR4. Consequently, we studied the FGFR4 gene as a candidate gene for genetic susceptibility to gallstone disease. We found that overexpression of FGFR4, especially the G-388R mutant of FGFR4, inhibits luciferase activity of CYP7A1 reporter in HepG2 cells, indicating that the G-388R mutant of FGFR4 may have greater inhibitory activity against bile acid biosynthesis. To investigate the association of FGFR4 polymorphism with gallstone disease, 117 patients with gallstone disease and 457 controls were genotyped for FGFR4 polymorphism G-388R by PCR-RFLP. Although the incidence of gallstone disease was not greater in patients with the FGFR4 RR genotype, the ratio of gallstone patients with acute cholecystitis in the FGFR4 RR genotype (42%) was significantly higher than that in other genotypes of FGFR4 (P = 0.019). In conclusion, the FGFR4 polymorphism is a genetic risk factor contributing to aggravation of gallstone disease.

Association between Fibroblast Growth Factor Receptor-4 Gene Polymorphism and Risk of Prostate Cancer: A Meta-Analysis

Objectives: To examine the association between fibroblast growth factor receptor-4 (FGFR-4) genetic polymorphisms (Gly-388Arg) and prostate cancer susceptibility. Methods: A comprehensive search was conducted to identify all case-control studies of FGFR-4 polymorphisms and prostate cancer risk. Statistical analysis was performed with the software program Stata, version 8.0, and Review Manage, version 4.2. Results: A total of 4 eligible studies, including 2,618 cases and 2,305 controls, relating the FGFR-4 polymorphism to the risk of prostate cancer were identified. The overall results indicated a significant association between FGFR-4 polymorphisms and prostate cancer. In the subgroup analysis by ethnicity, a significant association was found between European descent for recessive model (random effects OR 0.82, 95% CI 0.72–0.93) and co-dominant genetic model (random effects OR 0.83, 95% CI 0.68–1.00). Sensitivity analysis also found a significant association in the recessive (random effects OR 0.68, 95% CI 0.49–0.95) or the co-dominant (random effects OR 0.68, 95% CI 0.49–0.96) models. Egger’s test showed that publication bias was not present in any of the comparisons. Conclusions: Gly-388Arg polymorphism of FGFR-4 most likely contributes to susceptibility to prostate cancer, especially in men of European descent.

Abstract 1106: Fibroblast growth factor FGF19 signaling promotes prostate cancer progression

Abstract Aberrant fibroblast growth factor (FGF) signaling can promote tumor development by directly driving cancer cell proliferation and survival, and by supporting tumor angiogenesis. Multiple FGFs have been found upregulated in prostate cancers, including FGF1, FGF2, FGF6, FGF8 and FGF17, all of which can activate FGF receptor 4 (FGFR4). FGFR4 is overexpressed in prostate cancer (PCa) and positively associated with aggressive PCa. FGF19 is a distinct member of FGF family in that it predominantly binds to FGFR4 with high affinity. In this present study we aimed to study the role of FGF19 in human PCa progression, and to determine whether the targeted suppression of FGF19/FGFR4 signaling has potential therapeutic benefits in PCa. Our results demonstrated that FGF19 is upregulated in human PCa compared to normal prostate tissues. FGF19 is expressed in an autocrine manner by all tested PCa cell lines. Exogenous FGF19 stimulates PCa cell proliferation, anchorage-independent growth, adhesion and invasion in vitro. The mRNAs of FGF19 co-receptors αKlotho and ßKlotho are expressed in 97.5% and 27.5% of PCa clinical samples, respectively; but αKlotho is expressed in only 57% normal prostate tissues, ßKlotho is barely detected in normal prostate tissues. Suppression of FGF19 by short hairpin RNA (shRNA) targeting FGF19 gene inhibits PCa cell proliferation, adhesion and invasiveness in vitro. Immunoprecipitation and western blot assays showed that FGF19 stimulates phosphorylation of FGFR4, FRS2α, Erk1/2 and p-38 MAPK, as well as MEK1/2 in both PC3 and DU145. Our data also revealed that FGF19 induced the serine/threonine protein kinase Akt phosphorylation in PCa cells. Lentiviral shRNA delivery was used for stable FGFR4 gene silencing in PC3 and LNCaP cells. The targeted knockdown of FGFR4 in PC3 and LNCaP cells resulted in significantly inhibited cell proliferation, invasion and remarkably reduced Akt phosphorylation. The upregulated expression of activated Caspase 8 and activated Caspase 3 in FGFR4 knockdown cells indicates that FGFR4 signaling inhibits PCa cell apoptosis. To determine if FGFR4 suppression impacts prostate tumor growth and metastasis in vivo we generated a PCa orthotopic xenograft model in which PC3-sh-FGFR4 or PC3-shV cells are injected directly into the prostates of nude mice in each group. The results revealed that the FGFR4 suppression in PCa cells significantly inhibited tumorigenicity in this model (P=0.01). The primary tumor weight was decreased by 63% in sh-FRGR4 tumors (p&amp;lt;0.05) and the proportion of mice with lymph node metastasis was decreased from 93.3% to 50% (p&amp;lt;0.01). The Ki67 immunohistochemistry showed a significant decrease of Ki67 positive cell percentage in PC3-sh-FGFR4 xenograft tumors (19.85%) compared to control group (35.93%, p&amp;lt;0.001). Our data indicated that FGF19/FGFR4 signaling may be a promising target in PCa therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1106. doi:10.1158/1538-7445.AM2011-1106

Mechanism of regulation of FGFR4 gene expression by transcription factor Sp1 in human infant rhabdomyosarcomas

Objective To investigate the role of transcription factor Spl in the fibroblast growth factor receptor 4 （FGFR4） expression regulation and to obtain insight into the regulatory mechanisms governing FGFR4 gene regulation and overexpression in human infant rhabdomyosarcomas. Methods By using a series of cellular and molecular biology methods like transfection, Northern blotting, Western blotting etc. , the mRNA and protein expression of FGFR4 was detected. The regulatory effects of Spl on the FG- FR4 expression in the human rhabdomyosarcoma-derived cells were analyzed. Results Spl could be express highly in the human rhabdomyosarcoma-derived cells, and its binding to FGFR4 promoter could upregulate the FGFR4 expression, which was influenced by the methylation of FGFR4 promoter. Condusion Spl induces the expression of FGFR4 in human infant rhabdomyosareomas by directly binding of Spl with DNA cis elements in FGFR4 gene promoter region. Key words: Transcription factor; Gene expression ; Rhabdomyosarcomas

The Fibroblast Growth Factor Receptor-4 Arg388 Allele is Associated with Gastric Cancer Progression

Fibroblast growth factor receptor 4 (FGFR4) Gly388Arg polymorphism, located in the FGFR4 gene exon 9, was reported to be associated with malignant tumors prognosis; however, there has been no relevant research for gastric cancer. The purpose of this study was to investigate the clinical significance of FGFR4 Gly388Arg polymorphism as well as the mRNA expression of FGFR4 in patients with gastric cancer. The mRNA expression of FGFR4 in 103 gastric cancer tissues and corresponding normal tissues were measured by reverse transcription polymerase chain reaction (PCR) and real-time quantitative PCR. PCR-restriction fragment length polymorphism analysis was performed to detect the FGFR4 Gly388Arg in 103 gastric cancer tissues. In 57.3% of patients, homozygous or heterozygous Arg388 allele was present. FGFR4 expressions in mRNA levels were higher in gastric cancer tissues compared with those in relevant normal tissues. However, there is no significantly statistical difference compared with mRNA expression of FGFR4 in different genotypes. Associations between FGFR4 Gly388Arg polymorphism and overall survival exist in patients with gastric cancer (P = 0.046).The FGFR4 Arg allele (hazard risk (HR), 2.324; 95% confidence interval (CI), 1.054-4.125; P = 0.037) and TNM stage (HR, 5.516; 95% CI 3.658-7.409; P = 0.005) were independent prognostic factors in patients with gastric cancer. Based on this study, FGFR4 Arg388 genotype-a marker for gastric cancer progression-may predict prognosis of gastric cancer.