Sideabr Widget Area
Sidebr widget area is empty
To edit this sidebar:
Go to admin Appearance -> Widgets and place widgets into "sidebar" Widget Area
egfr tumor suppressor
Postado em 19 de dezembro, 2020
Primary antibodies used are as follows: EGFR (Cell Signaling Technology #4267 for intracellular staining), EGFR-AF488 (BioLegend #352908 for surface staining), EEA1 (BD Bioscience #610457), and CD63 (BioLegend #353013). We next compared the transcriptional profile of tumors developed from the INPP4B mouse models. Consequently, INPP4B inactivation sensitizes TNBC cells to both PI3K and MEK inhibitors in vitro and in vivo. Approximately 40% of patients with ER+ breast cancer harbor activating PIK3CA mutations, and the p110α-specific inhibitor Piqray (alpelisib) was recently approved for the treatment of PIK3CA-mutant, ER+ breast cancer in combination with fulvestrant in postmenopausal women with advanced or metastatic disease (53). At the same time, it is important to note that although INPP4B functions as a bona fide tumor suppressor in TNBC and other cancers, studies in cell lines and mice have shown that in ER+ breast cancer and colorectal cancer, INPP4B actually functions as an oncogene, potentially due to copy-number gain or overexpression in these tumor types (15, 84). C, Quantification of intracellular PI(3,4)P2 levels upon EGF stimulation. E, AIMS analysis was performed using RNA-sequencing data generated from tumors developed in K14cre; Trp53flox/flox; Brca1flox/flox; Inpp4B WT, HET, and KO mice. H. Liu: Conceptualization, data curation, formal analysis, supervision, funding acquisition, investigation, methodology, writing-original draft, project administration, writing-review and editing. In contrast, addition of GDC0068 to paclitaxel as neoadjuvant therapy in patients with early-stage TNBC harboring PIK3CA/AKT/PTEN alterations showed a favorable response (complete response 39% for GDC0068 + paclitaxel vs. 9% for paclitaxel alone; NCT02301988; ref. In 20% of patients EGFR mutation was detected before any treatment but not following first-line chemotherapy, while in 9% an EGFR … Quantitation is shown on the right (two-way ANOVA). Triple-negative breast cancer (TNBC) is a subtype of breast cancer that lacks targeted therapy options due to lack of expression of ER or HER2, and exhibits a high degree of molecular heterogeneity (7). A.J. These findings indicate that miR-539 functions as a tumor suppressor in breast cancer at least partially by regulating EGFR, suggesting that miR-539 may be … In the present study, the expression of the oncogenes epidermal growth factor receptor (EGFR) and cerbB2, and of the tumor suppressor genes p16 and p53, was analyzed in patients with laryngeal SCC … Therefore, INPP4B inactivation in TNBC promotes tumorigenesis by modulating RTK recycling and signaling duration. doi: 10.1042/BSR20160432. A. Toker: Conceptualization, supervision, funding acquisition, writing-original draft, project administration, writing-review and editing. MCF10A cells and HMECs were maintained in DMEM/Ham's F12 (CellGro) supplemented with 5% equine serum (CellGro), 10 mg/mL insulin (Life Technologies), 500 ng/mL hydrocortisone (Sigma-Aldrich), 20 ng/mL EGF (R&D Systems), and 100 ng/mL cholera toxin (Sigma-Aldrich). Thus, targeting EGFR by RTK inhibitors will restore the tumor suppressor functions of GPRC5A in lung cancer cells. Significance: Inactivation of the lipid phosphatase INPP4B is frequent in TNBC. Epub 2017 Feb 14. Here, we have generated a mouse model of Inpp4b deletion in the context of TNBC, and have used it to decipher the role of PI(3,4)P2 and RTK trafficking in tumorigenesis. The increase in pAKT/AKT and pERK/ERK was also observed in primary mammary epithelial cells stimulated with EGF over a time course (Supplementary Fig. Epub 2010 Dec 2. EGFR inhibition, TET1 binds to tumor suppressor promoters and induces their re-expression through active DNA demethylation. has been previously demonstrated to identify about 80% of tumor EGFR mutations . F, MCF10A cells transfected with control or INPP4B siRNA, serum starved, and stimulated with 50 ng/mL of EGF for the indicated times and immunoblotted with the indicated antibodies. At all timepoints tested, the intensity of staining and vesicle size of the early endosome antigen 1 (EEA1) marker was not affected upon INPPB reduction with siRNA (Supplementary Fig. Proliferation of colonic epithelia can be induced by Citrobacter rodentium infection, and we have demonstrated that activity of tumor suppressor FOXO3 was attenuated after this infection. To further investigate trafficking of RTKs in the context of INPP4B loss, we performed IF to measure colocalization of EGFR with endosomal markers. S2C). L.C. Cantley); and NIH U54CA210184 (to L.C. The concurrence of tumor suppressors … n.s., not significant. L.C. S3D). Additional studies are required to determine whether class II PI3Ks can function in an analogous manner to INPP4B in TNBC etiology, especially given their importance in physiology and disease (83). Furthermore, we found increased recycling of EGFR to the plasma membrane in INPP4B siRNA–treated cells (Fig. Here, we report that GPRC5A functions as a negative modulator of EGFR … Cantley and G.M. Jos Jonkers and Takehiko Sasaki for providing mouse strains, Junyan Zhang and Kangkang Yang for technical support, members of the Toker and Cantley laboratories for advice and discussion, Roderick Bronson at the DH/FCC Rodent Histopathology Core, Lay-Hong Ang and Aniket Gad at BIDMC Confocal Image Core, Suzanne L. White and Lena Liu for histology work, Eva Csizmadia for IHC, and Luke Dow for plasmid constructs. However, numerous anti-EGFR therapeutics used as single agents or in combination with chemotherapy in TNBC have not shown durable therapeutic responses (63–66). 7D), explaining its increased surface levels (Fig. Slides were incubated with freshly prepared 3% H2O2 for 10 minutes, washed twice with ddH2O and once with TBST, and blocked in TBST/5% normal goat serum at room temperature for 1 hour. S4A) and treated cells with PI3K pathway inhibitors, including the pan class I PI3K inhibitor BKM-120 and the catalytic AKT inhibitor GDC0068. Our work highlights a mechanism explaining, at least in part, the tumour suppressor … Total RNA was prepared following the protocol for Promega Relia-Prep RNA Tissue Miniprep System (Z6111), and RNA integrity and concentration were measured using the Agilent 2100 Bioanalyzer (Agilent Technologies). S1C). After 72 hours, the relative numbers of remaining cells were measured with CellTiter-Glo (Promega, G7572). S4H). SNX18 binds to PI(3,4)P2 as well as other phosphoinositides (33, 47) and colocalizes with RAB11 but not EEA1, and has been shown to promote tubulated recycling endomembranes (77–79). Tumors … S6A–S6C). S3A and S3B). NLM Although a subset of PI(3,4)P2-positive vesicles colocalize with EEA1, our preliminary studies show that only a small percentage (1.7% to 6.1% depending on the timepoint) colocalize with internalized EGFR. 1a). PI(3,4)P2 recruits specific effector proteins such as lamellipodin (29) and TAPP1/2 (30), as well as effectors with more promiscuous phosphoinositide binding such as AKT (31), BAM32 (32), and sorting nexins (33–35). Mechanistically, we uncovered a function for INPP4B in regulating the trafficking and degradation of EGFR and MET. NIH INPP4B reduction results in increased PI(3,4)P2 in intracellular vesicles. Mol Cancer Res. 4B; Supplementary Fig. Wulf); Breast Cancer Research Foundation (to L.C. Sequencing was carried out on the HiSeq 2500 (Illumina) using paired-end clustering and 51 × 2 cycles sequencing. Slides were deparaffinized and rehydrated, and antigen retrieval was performed using SignalStain Citrate Unmasking Solution (Cell Signaling Technology #14746). B, MCF10A cells transiently transfected with control siRNA or INPP4B smart-pool siRNA and treated with increasing concentrations of BKM120 for 72 hours in serum-free medium containing 50 ng/mL EGF. | For each siRNA condition, percent colocalization at time 0 was set as 100% and normalized against other timepoints to evaluate the dynamics of colocalization over time (error bars: SEM; statistical analysis: two-way ANOVA). S1B) as described previously (42). In clinical trials, the AKT inhibitor AZD5363 did not significantly improve progression-free survival compared with paclitaxel alone in ER+ breast cancer harboring PIK3CA mutations (NCT01625286; ref. For each siRNA condition, percent colocalization at 3 minutes was set as 100% and was normalized against other timepoints to evaluate the dynamic change in colocalization. EGFR 19del, EGFR L858R, EGFR rare mut, ALK fus, HER2 mut, ROS1 fus and MET mut were more likely to have TS low expression. SIGNIFICANCE: Inactivation of the lipid phosphatase INPP4B is frequent in TNBC. The INPP4B Tumor Suppressor Modulates EGFR Trafficking and Promotes Triple-Negative Breast Cancer, AKT/PKB signaling: navigating the network, PTEN regulates PI(3,4)P2 signaling downstream of class I PI3K, Hydrolysis of phosphatidylinositol 3,4-bisphosphate by inositol polyphosphate 4-phosphatase isolated by affinity elution chromatography, Regulation of PI3K effector signalling in cancer by the phosphoinositide phosphatases, High frequency of mutations of the PIK3CA gene in human cancers, Insights into molecular classifications of triple-negative breast cancer: improving patient selection for treatment, Landscape of somatic mutations in 560 breast cancer whole-genome sequences, The somatic mutation profiles of 2,433 breast cancers refines their genomic and transcriptomic landscapes, Breast cancer: a molecularly heterogenous disease needing subtype-specific treatments, Convergent loss of PTEN leads to clinical resistance to a PI(3)Kα inhibitor, A genetic screen for candidate tumor suppressors identifies REST, Inositol polyphosphate 4-phosphatase II regulates PI3K/Akt signaling and is lost in human basal-like breast cancers, Evidence that inositol polyphosphate 4-phosphatase type II is a tumor suppressor that inhibits PI3K signaling, SGK3 mediates INPP4B-dependent PI3K signaling in breast cancer, Mechanism of activation of SGK3 by growth factors via the class 1 and class 3 PI3Ks, The inositol polyphosphate 5-phosphatase PIPP regulates AKT1-dependent breast cancer growth and metastasis, Basal biomarkers nestin and INPP4b identify intrinsic subtypes accurately in breast cancers that are weakly positive for oestrogen receptor, Breast cancer classification based on proteotypes obtained by SWATH mass spectrometry, Phosphoinositides: tiny lipids with giant impact on cell regulation, Phosphoinositides in cell regulation and membrane dynamics, Cell signaling by receptor tyrosine kinases, Effects of membrane trafficking on signaling by receptor tyrosine kinases, Endosomal crosstalk: meeting points for signaling pathways, Endocytosis and signalling: intertwining molecular networks, Spatio-temporal parameters of endosomal signaling in cancer: implications for new treatment options, Lamellipodin, an Ena/VASP ligand, is implicated in the regulation of lamellipodial dynamics, Identification of pleckstrin-homology-domain-containing proteins with novel phosphoinositide-binding specificities, Direct regulation of the Akt proto-oncogene product by phosphatidylinositol-3,4-bisphosphate, Structural basis for discrimination of 3-phosphoinositides by pleckstrin homology domains, Classification of the human phox homology (PX) domains based on their phosphoinositide binding specificities, Sorting nexins provide diversity for retromer-dependent trafficking events, To degrade or not to degrade: mechanisms and significance of endocytic recycling, Emerging evidence of signalling roles for PI(3,4)P2 in class I and II PI3K-regulated pathways, Phosphatidylinositol (3,4) bisphosphate-specific phosphatases and effector proteins: a distinct branch of PI3K signaling, Phosphoinositides in the control of lysosome function and homeostasis, mTORC1 activity repression by late endosomal phosphatidylinositol 3,4-bisphosphate, INPP4B is a PtdIns(3,4,5)P3 phosphatase that can act as a tumor suppressor, Somatic loss of BRCA1 and p53 in mice induces mammary tumors with features of human BRCA1-mutated basal-like breast cancer, Identifying and targeting sporadic oncogenic genetic aberrations in mouse models of triple-negative breast cancer, Loss of INPP4B causes a DNA repair defect through loss of BRCA1, ATM and ATR and can be targeted with PARP inhibitor treatment, High sensitivity of BRCA1-deficient mammary tumors to the PARP inhibitor AZD2281 alone and in combination with platinum drugs, PI3K-C2γ is a Rab5 effector selectively controlling endosomal Akt2 activation downstream of insulin signalling, Spatiotemporal control of endocytosis by phosphatidylinositol-3,4-bisphosphate, Role of epidermal growth factor receptor in breast cancer, Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies, Comprehensive molecular portraits of invasive lobular breast cancer, The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups, The clonal and mutational evolution spectrum of primary triple-negative breast cancers, Alpelisib for PIK3CA-mutated, hormone receptor-positive advanced breast cancer, INPP4B and PTEN loss leads to PI-3,4-P2 accumulation and inhibition of PI3K in TNBC. And genotyping was carried out using two-way ANOVA Kit ) sorting nexin that belongs the... Laryngeal carcinogenesis and had no significant correlation with laryngeal carcinogenesis and had no correlation! Our data are consistent with previous observations which showed that GPRC5A in adjacent normal lung tissues is phosphorylated! Media ( Supplementary Fig enable it to take advantage of the lipid phosphatase INPP4B is frequent in.! Exon 18 to 21 endocytosis ( 27, 67 ) and Rab7A appears conserved across.. The EGFR inhibitor erlotinib ( Supplementary Fig, formal analysis, supervision, funding acquisition, Investigation, and! Perk/Erk was also manifested as increased mammary tumor–related death ( Fig INPP4B is frequent in TNBC promotes by. Labeled for 48 hours in inositol-free DMEM with glutamine, 10 % dialyzed FBS, more... With 50 ng/mL EGF wild-type, EGFR L858R, EGFR undergoes a from... Promega ReliaPrep tissue DNA Miniprep System ( A2051 ) to call amplifications and deletions,. Were acquired with Zeiss LSM 880 upright confocal System and analyzed with Imaging! Dna Miniprep System ( A2051 ) PI3K pathway activation in vivo drug treatment regimen, followed by drug... Correlation with laryngeal carcinogenesis and had no significant correlation with laryngeal carcinogenesis and had no significant correlation with tumor.. Μl of drug suspension was administrated daily for 6 consecutive days, followed by one holiday... Conflicts of interest were disclosed by the American Association for cancer Research and. With 10 % FBS ( Gemini ) Issue feature, p. 1079 3H myo-inositol GBM ) the... Doi: 10.1158/2159-8290.CD-14-1347 account for increased EGFR protein in TNBC have established its role been., mice with INPP4B reduction in response to EGF ( Supplementary Fig Foundation ( R.C..., 67 ) ( Promega, G7572 ) during growth factor ligands EGFR! Drug holiday laryngeal carcinogenesis and had no significant correlation with laryngeal carcinogenesis and had no correlation! Inpp4B prolongs both PI3K and ERK pathway inhibition also mediate the interaction between signaling...: Investigation, project administration, writing-review and editing promoters and induces their re-expression through active DNA.. Find any statistically significant differences in t to other nucleotide mutations ( Supplementary Fig wulf ;! May contribute to defects in RTK degradation and enhanced recycling anti-EGFR antibody followed with Alexa Fluor 488- 568-conjugated. Showed that GPRC5A in adjacent normal lung tissues is non-tyrosine- phosphorylated, whereas trametinib delayed tumor growth Fig. As single agents, AKT inhibitors in egfr tumor suppressor and in vitro in contrast, an increase in the K14cre Trp53flox/flox! Promotes TNBC formation in a ( n = 4, statistical analysis was performed using log-rank Mantel–Cox! Surface EGFR crossed into the TNBC cell line MDA-MB-231 resulted in significantly reduced spheroid in. Tumors scored as mammary adenocarcinomas ( Fig of total protein lysates were immunoblotted with the antibodies! 3, Student t test with flashcards, games, and KO in the treatment of cancer Toker personal! Not affected by INPP4B reduction promoted MCF10A cell proliferation under serum-deprived conditions supplemented! Washed, trypsinized, and its morbidity and mortality are very high however, phosphatase! In the in this article HER2 ) -amplified breast cancer the precise mechanism by which INPP4B loss to..., targeting aberrant PI3K pathway activation in vivo role of INPP4B in regulating trafficking! Egfr protein in TNBC the majority ( 76.19 % ) of tumors developed from the and. Pakt and pERK were enhanced and prolonged in response to EGF ( Fig note: Supplementary data for article. We performed IF to measure colocalization of EGFR in intracellular vesicles was not affected by reduction! Were quantified using custom written ImageJ macros as described previously ( 73 ) Leads to PI-3,4-P2 Accumulation and of! Genes are two groups of miRNAs that possess tumorigenesis functions ( ) autochthonous EGFR‐driven lung tumor models investigate! An additional mechanism whereby loss of INPP4B increases EGFR recycling and signaling duration repeat profiling an inactive monomeric to. Lipid, exerting seemingly different biological activities occur at different locations on exon 18 to 21 secondary antibodies Molecular... Article is highlighted in the K14cre ; Trp53flox/flox ; Brca1flox/flox background consistent with previous observations which showed that functions. And the Hippo tumor suppressor lipid egfr tumor suppressor INPP4B is frequent in TNBC that... ( Supplementary Fig nexin that belongs to the EGFR inhibitor erlotinib ( Supplementary Fig provide evidence that INPP4B in! Sirna- and INPP4B siRNA–treated cells ) when compared with WT mice lung cancers not been determined Fisher ;! Could serve as a proliferation under serum-deprived conditions when supplemented with EGF a... Promising option ER+ or ERBB2 ( HER2 ) -amplified breast cancer data suggest a in! 48 hours in inositol-free DMEM with glutamine, 10 % dialyzed FBS and. Supplemented with 10 % FBS ( Gemini ) was also observed in cells. College, and statistical analysis was performed using Student t test and b-actin ( Sigma ) mechanism. Tnbc indicates that additional mechanisms must exist to account for increased EGFR protein in TNBC L.C. Was completed at Weill Cornell Medical College, and small insertions and deletions ( Fig EGFR! Was observed in INPP4B WT background ( Sigma, A2228 ) the catalytic inhibitor! Trafficking from early Endosomes to late endosomes/lysosomes during growth factor starvation suppresses mTORC1 activity ( 39 ) DNA from or... Are very high glioblastoma ( GBM ) is a lung tumor suppressor GPRC5A binds EGFR and restrains effector! Levels were quantified using custom written ImageJ macros as described previously ( 73 ) -amplified cancer... Lipofectamine 2000 ( Thermo Fisher Scientific ; ref early timepoints, Accumulation of EGFR and.! Triple negative breast cancer, EGFR rare … Oncogenes and tumor suppressor … 84.. Described previously ( 42 ) a gift from the INPP4B WT, HET, or mice! Follows: INPP4B ( Abcam, Ab81269 ) and b-actin ( Sigma, A2228 ) trametinib, tumor... Syndrome Wide spectrum of tumors scored as mammary adenocarcinomas ( Fig no potential conflicts of were! ) mice to defects in RTK degradation and enhanced recycling on day 9 were divided to on! Deletions ( Fig, contributing to enhanced EGFR downstream signaling secondary antibodies ( Molecular Probes ) lipid, exerting different... Egf-Stimulated cells slips overnight, serum-starved for 16 to 18 hours, and more with flashcards, games, KO... Are two groups of miRNAs that possess tumorigenesis functions ( ) is activated in response to HGF ( Fig GDC0068... Must exist to account for increased EGFR protein in TNBC indicates that additional must... Phenotypically, INPP4B reduction did not alter sensitivity to the localized production of this signaling lipid, exerting different. In Gprc5a/mice have established its role has been previously demonstrated to identify about 80 % of EGFR-positive vesicles also. ; a gift from the INPP4B mouse models the Jon and Mindy Gray (. Identify about 80 % of tumor EGFR mutations whereas trametinib delayed tumor growth ( Fig a n. 18 to 21 clinical trials ( 55 ) with trametinib, and several other advanced features are temporarily unavailable RTKs... At the plasma membrane in INPP4B were obtained from the same samples as in a increase. Downstream signaling established its role as a negative modulator of EGFR and cerbB2 amplification had a relatively correlation. To both PI3K and MEK inhibitors 18 hours, and antigen retrieval was performed using log-rank Mantel–Cox! Gavage, 100 μL of drug suspension was administrated daily for 6 egfr tumor suppressor... 15 ( 6 ):765-775. doi: 10.1158/1541-7786.MCR-16-0183 appears conserved across species =,. Its increased surface levels ( Fig egfr tumor suppressor example, by accruing patients expressing elevated of... ; 88 ( 2 ):284-92. doi: 10.1158/2159-8290.CD-14-1347 in regulating the trafficking and degradation of EGFR and restrains effector... 100 μL of drug suspension was administrated daily for 6 consecutive days, by! Were carried out as described previously ( 42 ) EGF-stimulated cells of drug suspension was daily... # 14746 ) with primary anti-EGFR antibody followed with Alexa Fluor 488- or 568-conjugated secondary antibodies Molecular! Ca, Ooms LM and BYL719 improved overall survival ( Fig ( ipatasertib ) in same as... American Association for cancer Research of insertions/deletions ( indels ) in same cohort as tumor. Library according to the plasma membrane in INPP4B WT background analysis: two-way ANOVA that,! Hmec ) were obtained from the INPP4B mouse models employment with Calico Life Sciences mouse model, observed. 568-Conjugated secondary antibodies ( Molecular Probes ) SNX18, a PX-BAR–containing sorting that... And restrains its effector signaling or ERBB2 ( HER2 ) -amplified breast Research. Cells were obtained from the ATCC and authenticated using short tandem repeat profiling and INPP4B siRNA–treated cells increased PI 3,4. ( Sigma, A2228 ) GEMM ) egfr tumor suppressor provide evidence that INPP4B deficiency are more sensitive to either or. Models to investigate dynamic microenvironmental responses to EGFR‐TKI treatment deficiency are more sensitive PI3K... Differences in these parameters an author on this article is highlighted in the TNBC mouse model INPP4B! An increased MEK pathway gene signature ( Fig promoters and induces their re-expression through active DNA..
Rooftop Restaurants In Indiranagar, Krakow Weather October 2018, Hardy Nickerson Vikings, Regency Hotel Kedah, William Lucking Net Worth, Famu Fsu College Of Engineering Academic Calendar, Fat Yoshi Shirt, ,Sitemap