Search tips
Search criteria

Results 1-20 (20)

Clipboard (0)

Select a Filter Below

more »
Year of Publication
1.  Loss of PTEN expression is associated with poor prognosis in patients with intraductal papillary mucinous neoplasms of the pancreas 
Previously, we reported PIK3CA gene mutations in high-grade intraductal papillary mucinous neoplasms (IPMN). However, the contribution of phosphatidylinositol-3 kinase pathway (PI3K) dysregulation to pancreatic carcinogenesis is not fully understood and its prognostic value unknown. We investigated the dysregulation of the PI3K signaling pathway in IPMN and its clinical implication.
Experimental Design
Thirty-six IPMN specimens were examined by novel mutant-enriched methods for hot-spot mutations in the PIK3CA and AKT1 genes. PIK3CA and AKT1 gene amplifications and loss of heterozygosity (LOH) at the PTEN locus were also evaluated. Additionally, the expression levels of PDPK1/PDK1, PTEN and Ki67 were analyzed by immunohistochemistry.
Three cases carrying the E17K mutation in the AKT1 gene and one case harboring the H1047R mutation in the PIK3CA gene were detected among the 36 cases. PDK1 was significantly overexpressed in the high-grade IPMN vs. low-grade IPMN (p = 0.034) and in pancreatic and intestinal-type of IPMN vs. gastric-type of IPMN (p = 0.020). Loss of PTEN expression was strongly associated with presence of invasive carcinoma and poor survival in these IPMN patients (p = 0.014).
This is the first report of AKT1 mutations in IPMN. Our data indicate that oncogenic activation of the PI3K pathway can contribute to the progression of IPMN, in particular loss of PTEN expression. This finding suggests the potential employment of PI3K pathway-targeted therapies for IPMN patients. The incorporation of PTEN expression status in making surgical decisions may also benefit IPMN patients and should warrant further investigation.
PMCID: PMC3915026  PMID: 24132918
PI3K-pathway; AKT1; PTEN; IPMN; mutant-enriched sequencing method
2.  Metformin and erlotinib synergize to inhibit basal breast cancer 
Oncotarget  2014;5(21):10503-10517.
Basal-like breast cancers (BBCs) are enriched for increased EGFR expression and decreased expression of PTEN. We found that treatment with metformin and erlotinib synergistically induced apoptosis in a subset of BBC cell lines. The drug combination led to enhanced reduction of EGFR, AKT, S6 and 4EBP1 phosphorylation, as well as prevented colony formation and inhibited mammosphere outgrowth. Our data with other compounds suggested that biguanides combined with EGFR inhibitors have the potential to outperform other targeted drug combinations and could be employed in other breast cancer subtypes, as well as other tumor types, with activated EGFR and PI3K signaling. Analysis of BBC cell line alterations led to the hypothesis that loss of PTEN sensitized cells to the drug combination which was confirmed using isogenic cell line models with and without PTEN expression. Combined metformin and erlotinib led to partial regression of PTEN-null and EGFR-amplified xenografted MDA-MB-468 BBC tumors with evidence of significant apoptosis, reduction of EGFR and AKT signaling, and lack of altered plasma insulin levels. Combined treatment also inhibited xenografted PTEN null HCC-70 BBC cells. Measurement of trough plasma drug levels in xenografted mice and a separately performed pharmacokinetics modeling study support possible clinical translation.
PMCID: PMC4279389  PMID: 25361177
breast cancer; metformin; erlotinib; PTEN; EGFR
3.  RFP-mediated ubiquitination of PTEN modulates its effect on AKT activation 
Cell Research  2013;23(4):552-564.
The PTEN tumor suppressor is a lipid phosphatase that has a central role in regulating the phosphatidylinositol-3-kinase (PI3K) signal transduction cascade. Nevertheless, the mechanism by which the PTEN activity is regulated in cells needs further elucidation. Although previous studies have shown that ubiquitination of PTEN can modulate its stability and subcellular localization, the role of ubiquitination in the most critical aspect of PTEN function, its phosphatase activity, has not been fully addressed. Here, we identify a novel E3 ubiquitin ligase of PTEN, Ret finger protein (RFP), that is able to promote atypical polyubiquitinations of PTEN. These ubiquitinations do not lead to PTEN instability or relocalization, but rather significantly inhibit PTEN phosphatase activity and therefore modulate its ability to regulate the PI3K signal transduction cascade. Indeed, RFP overexpression relieves PTEN-mediated inhibitory effects on AKT activation; in contrast, RNAi-mediated knockdown of endogenous RFP enhances the ability of PTEN to suppress AKT activation. Moreover, RFP-mediated ubiquitination of PTEN inhibits PTEN-dependent activation of TRAIL expression and also suppresses its ability to induce apoptosis. Our findings demonstrate a crucial role of RFP-mediated ubiquitination in controlling PTEN activity.
PMCID: PMC3616435  PMID: 23419514
PTEN; RFP; ubiquitination; AKT; phosphorylation
4.  A secreted PTEN phosphatase that enters cells to alter signaling and survival 
Science (New York, N.Y.)  2013;341(6144):399-402.
Phosphatase and Tensin Homologue on chromosome Ten (PTEN) is a tumor suppressor and an antagonist of the phosphoinositide-3 kinase (PI3K) pathway. We identified a 576-amino acid translational variant of PTEN, termed PTEN-Long, that arises from an alternative translation start site 519 bp upstream of the ATG initiation sequence, adding 173 N-terminal amino acids to the normal PTEN open reading frame. PTEN-Long is a membrane permeable lipid phosphatase that is secreted from cells and can enter other cells. As an exogenous agent, PTEN-Long antagonized PI3K signaling and induced tumor cell death in vitro and in vivo. By providing a means to restore a functional tumor suppressor protein to tumor cells, PTEN-Long may have therapeutic uses.
PMCID: PMC3935617  PMID: 23744781
5.  Alterations of EGFR, p53 and PTEN that mimic changes found in basal-like breast cancer promote transformation of human mammary epithelial cells 
Cancer Biology & Therapy  2013;14(3):246-253.
Breast cancer can be classified into different molecular subtypes with varying clinical and pathological characteristics. The basal-like breast cancer subtype represents one of the most aggressive and lethal types of breast cancer, and due to poor mechanistic understanding, it lacks targeted therapy. Many basal-like breast cancer patient samples display alterations of established drivers of cancer development, including elevated expression of EGFR, p53 inactivating mutations and loss of expression of the tumor suppressor PTEN; however, their contribution to human basal-like breast cancer pathogenesis remains ill-defined. Using non-transformed human mammary epithelial cells, we set out to determine whether altering EGFR, p53 and PTEN in different combinations could contribute to basal-like breast cancer progression through transformation of cells. Altering PTEN in combination with either p53 or EGFR in contrast to any of the single alterations caused increased growth of transformed colonies in soft agar. Concomitantly modifying all three genes led to the highest rate of cellular proliferation and the greatest degree of anchorage-independent colony formation. Results from our effort to engineer a model of BBC expressing alterations of EGFR, p53 and PTEN suggest that these changes are cooperative and likely play a causal role in basal-like breast cancer pathogenesis. Consideration should be given to targeting EGFR and restoring p53 and PTEN signaling simultaneously as a strategy for treatment of this subtype of breast cancer.
PMCID: PMC3595307  PMID: 23291982
basal-like breast cancer; EGFR; PTEN; p53; dominant-negative; MCF10A; oncogene; tumor suppressor; transformation; soft agar
Molecular cell  2010;40(6):877-892.
While the small GTPase Rac1 and its effectors are well-established mediators of mitogenic and motile signaling by tyrosine-kinase receptors and have been implicated in breast tumorigenesis, little is known regarding the exchange factors (Rac-GEFs) that mediate ErbB receptor responses. Here we identify the PIP3-Gβγ-dependent Rac-GEF P-Rex1 as an essential mediator of Rac1 activation, motility, cell growth, and tumorigenesis driven by ErbB receptors in breast cancer cells. Notably, activation of P-Rex1 in breast cancer cells requires the convergence of inputs from ErbB receptors and a Gβγ- and PI3Kγ-dependent pathway. Moreover, we identified the GPCR CXCR4 as a crucial mediator of P-Rex1/Rac1 activation in response to ErbB ligands. P-Rex1 is highly overexpressed in human breast cancers and their derived cell lines, particularly those with high ErbB2 and ER expression. In addition to the prognostic and therapeutic implications, our findings reveal an ErbB effector pathway that is crucial for breast cancer progression.
PMCID: PMC3038344  PMID: 21172654
P-Rex1; Rac-GEF; Rac1; ErbB receptors; PI3Kγ; Gβγ subunits; breast cancer
7.  Reduction of Pten dose leads to neoplastic development in multiple organs of PtenshRNA mice 
Cancer Biology & Therapy  2010;10(11):1194-1200.
To address the impact of partial reduction of Pten on tumor initiation, we generated PtenshRNA mice, in which PTEN expression was reduced below normal levels in various tissues. PtenshRNA mice frequently developed lymphoid and prostatic hyperplasia, splenomegaly and sebaceous adenomas. Our observations support the notion that partial reduction of the dose of Pten with shRNA is sufficient to induce neoplastic disease in multiple organ systems.
PMCID: PMC3018670  PMID: 20980828
pten; shRNA; RNAi; neoplasia; haploinsufficiency; tumor suppressor
8.  Reduction of Pten dose leads to neoplastic development in multiple organs of PtenshRNA mice 
Cancer biology & therapy  2010;10(11):1194-1200.
To address the impact of partial reduction of Pten on tumor initiation, we generated PtenshRNA mice, in which PTEN expression was reduced below normal levels in various tissues. PtenshRNA mice frequently developed lymphoid and prostatic hyperplasia, splenomegaly and sebaceous adenomas. Our observations support the notion that partial reduction of the dose of Pten with shRNA is sufficient to induce neoplastic disease in multiple organ systems.
PMCID: PMC3018670  PMID: 20980828
pten; shRNA; RNAi; neoplasia; haploinsufficiency; tumor suppressor
9.  Recurrent gross mutations of the PTEN tumor suppressor gene in breast cancers with deficient DSB repair 
Nature genetics  2007;40(1):102-107.
Basal-like breast cancer (BBC) is a subtype of breast cancer with poor prognosis1–3. Inherited mutations of BRCA1, a cancer susceptibility gene involved in double-strand DNA break (DSB) repair, lead to breast cancers that are nearly always of the BBC subtype3–5; however, the precise molecular lesions and oncogenic consequences of BRCA1 dysfunction are poorly understood. Here we show that heterozygous inactivation of the tumor suppressor gene Pten leads to the formation of basal-like mammary tumors in mice, and that loss of PTEN expression is significantly associated with the BBC subtype in human sporadic and BRCA1-associated hereditary breast cancers. In addition, we identify frequent gross PTEN mutations, involving intragenic chromosome breaks, inversions, deletions and micro copy number aberrations, specifically in BRCA1-deficient tumors. These data provide an example of a specific and recurrent oncogenic consequence of BRCA1-dependent dysfunction in DNA repair and provide insight into the pathogenesis of BBC with therapeutic implications. These findings also argue that obtaining an accurate census of genes mutated in cancer will require a systematic examination for gross gene rearrangements, particularly in tumors with deficient DSB repair.
PMCID: PMC3018354  PMID: 18066063
10.  Activation of the PI3K Pathway in Cancer through Inhibition of PTEN by Exchange Factor P-REX2a 
Science (New York, N.Y.)  2009;325(5945):1261-1265.
PTEN (Phosphatase and tensin homolog on chromosome ten) is a tumor suppressor whose cellular regulation remains incompletely understood. We identified Phosphatidylinositol-3,4,5-trisphosphate RAC Exchanger 2a (P-REX2a) as a PTEN-interacting protein. P-REX2a mRNA was more abundant in cancer, and significantly increased in tumors with wild type PTEN that expressed an activated mutant of PIK3CA encoding the p110 subunit of phosphoinositide 3-kinase-α (PI3Kα). P-REX2a inhibited PTEN lipid phosphatase activity and stimulated the PI3K pathway only in the presence of PTEN. P-REX2a stimulated cell growth and cooperated with a PIK3CA mutant to promote growth factor-independent proliferation and transformation. Depletion of P-REX2a reduced amounts of phosphorylated AKT and growth in cell lines with intact PTEN. Thus P-REX2a is a component of the PI3K pathway that can antagonize PTEN in cancer cells.
PMCID: PMC2936784  PMID: 19729658
11.  BAF180 is a critical regulator of p21 induction and a tumor suppressor mutated in breast cancer 
Cancer research  2008;68(6):1667-1674.
Screening for tumor suppressor genes in breast cancer revealed multiple truncating mutations of PB1, which encodes the BAF180 subunit of the PBAF chromatin remodeling complex. Mutation was associated with loss of heterozygosity of the wild type allele. BAF180 complementation of BAF180-mutant tumor cells caused G1 arrest that was dependent upon increased expression of the cyclin/cdk inhibitor p21/WAF1/Cip1. Endogenous wild type BAF180 bound to the p21 promoter and was required for proper p21 expression and G1 arrest after TGF-β and γ-radiation treatment. BAF180 thus functions on two tumor suppressor signaling pathways as a physiological mediator of p21 expression. We conclude that BAF180 suppresses tumorigenesis at least in part through its ability to regulate p21.
PMCID: PMC2915562  PMID: 18339845
BAF180; PB1; p21/WAF1/CIP1; tumor suppressor; breast cancer
12.  PDK1 potentiates upstream lesions on the PI3K pathway in breast carcinoma 
Cancer research  2009;69(15):6299-6306.
Lesions of ERBB2, PTEN, and PIK3CA activate the PI3K pathway during cancer development by increasing levels of phosphatidylinositol-3,4,5-triphosphate (PIP3). 3-phosphoinositide-dependent kinase 1 (PDK1) is the first node of the PI3K signal output and is required for activation of AKT. PIP3 recruits PDK1 and AKT to the cell membrane through interactions with their PH domains, allowing PDK1 to activate AKT by phosphorylating it at residue threonine 308. We show that total PDK1 protein and mRNA was over-expressed in a majority of human breast cancers and that 21% of tumors had five or more copies of the gene encoding PDK1, PDPK1. We found that increased PDPK1 copy number was associated with upstream pathway lesions (ERBB2 amplification, PTEN loss, or PIK3CA mutation), as well as patient survival. Examination of an independent set of breast cancers and tumor cell lines derived from multiple forms of human cancers also found increased PDK1 protein levels associated with such upstream pathway lesions. In human mammary cells, PDK1 enhanced the ability of upstream lesions to signal to AKT, stimulate cell growth and migration, and rendered cells more resistant to PDK1 and PI3K inhibition. After orthotopic transplantation, PDK1 overexpression was not oncogenic but dramatically enhanced the ability of ERBB2 to form tumors. Our studies argue that PDK1 overexpression and increased PDPK1 copy number are common occurrences in cancer that potentiate the oncogenic effect of upstream lesions on the PI3K pathway. Therefore, we conclude that alteration of PDK1 is a critical component of oncogenic PI3K signaling in breast cancer.
PMCID: PMC2727605  PMID: 19602588
PDK1; PI3K; ERBB2; PTEN; breast
13.  Distinct IL-2 Receptor Signaling Pattern in CD4+CD25+ Regulatory T Cells1 
Despite expression of the high-affinity IL-2R, CD4+CD25+ regulatory T cells (Tregs) are hypoproliferative upon IL-2R stimulation in vitro. However the mechanisms by which CD4+CD25+ T cells respond to IL-2 signals are undefined. In this report, we examine the cellular and molecular responses of CD4+CD25+ Tregs to IL-2. IL-2R stimulation results in a G1 cell cycle arrest, cellular enlargement and increased cellular survival of CD4+CD25+ T cells. We find a distinct pattern of IL-2R signaling in which the Janus kinase/STAT pathway remains intact, whereas IL-2 does not activate downstream targets of phosphatidylinositol 3-kinase. Negative regulation of phosphatidylinositol 3-kinase signaling and IL-2-mediated proliferation of CD4+CD25+ T cells is inversely associated with expression of the phosphatase and tensin homologue deleted on chromosome 10, PTEN.
PMCID: PMC2842445  PMID: 15100267
14.  PTEN Loss Promotes Mitochondrially Dependent Type II Fas-Induced Apoptosis via PEA-15▿  
Molecular and Cellular Biology  2008;29(5):1222-1234.
Two distinct biochemical signals are delivered by the CD95/Fas death receptor. The molecular basis for the differential mitochondrially independent (type I) and mitochondrially dependent (type II) Fas apoptosis pathways is unknown. By analyzing 24 Fas-sensitive tumor lines, we now demonstrate that expression/activity of the PTEN tumor suppressor strongly correlates with the distinct Fas signals. PTEN loss-of-function and gain-of-function studies demonstrate the ability to interconvert between type I and type II Fas pathways. Importantly, from analyses of Bcl-2 transgenic Pten+/− mice, Pten haploinsufficiency converts Fas-induced apoptosis from a Bcl-2-independent to a Bcl-2-sensitive response in primary thymocytes and activated T lymphocytes. We further show that PTEN influences Fas signaling, at least in part, by regulating PEA-15 phosphorylation and activity that, in turn, regulate the ability of Bcl-2 to suppress Fas-induced apoptosis. Thus, PTEN is a key molecular rheostat that determines whether a cell dies by a mitochondrially independent type I versus a mitochondrially dependent type II apoptotic pathway upon Fas stimulation.
PMCID: PMC2643821  PMID: 19103758
15.  Mutational loss of PTEN induces resistance to NOTCH1 inhibition in T-cell leukemia 
Nature medicine  2007;13(10):1203-1210.
Gain-of-function mutations in NOTCH1 are common in T-cell lymphoblastic leukemias (T-ALL), making this receptor a promising target for drugs such as γ-secretase inhibitors, which block a proteolytic cleavage required for NOTCH1 activation. However, the enthusiasm for these therapies has been tempered by tumor resistance and the paucity of information on the oncogenic programs regulated by oncogenic NOTCH1. Here we show that NOTCH1 regulates PTEN expression and the activity of the PI3K-AKT signaling pathway in normal and leukemic T cells. Notch signaling and the PI3K-AKT pathway synergize in vivo in a Drosophila model of Notch-induced tumorigenesis, and mutational loss of PTEN is associated with resistance to NOTCH1 inhibition in human T-ALL. Overall, these findings identify the transcriptional control of PTEN and the regulation of the PI3K/AKT pathway as key elements of the leukemogenic program activated by NOTCH1 and provide the basis for the design of new therapeutic strategies for T-ALL.
PMCID: PMC2600418  PMID: 17873882
16.  Methylation of the PTEN promoter defines low-grade gliomas and secondary glioblastoma 
Neuro-Oncology  2007;9(3):271-279.
Glioblastoma multiforme (GBM) can present as either de novo or secondary tumors arising from previously diagnosed low-grade gliomas. Although these tumor types are phenotypically indistinguishable, de novo and secondary GBMs are associated with distinct genetic characteristics. PTEN mutations, which result in activation of the phosphoinositide 3-kinase (PI3K) signal transduction pathway, are frequent in de novo but not in secondary GBMs or their antecedent low-grade tumors. Results we present here show that grade II astrocytomas, oligodendrogliomas, and oligoastrocytomas commonly display methylation of the PTEN promoter, a finding that is absent in nontumor brain specimens and rare in de novo GBMs. Methylation of the PTEN promoter correlates with protein kinase B (PKB/Akt) phosphorylation, reflecting functional activation of the PI3K pathway. Our results also demonstrate frequent methylation of the PTEN promoter in grade III astrocytomas and secondary GBMs, consistent with the hypothesis that these tumors arise from lower grade precursors. PTEN methylation is rare in de novo GBMs and is mutually exclusive with PTEN mutations. We conclude that methylation of the PTEN promoter may represent an alternate mechanism by which PI3K signaling is increased in grade II and III gliomas as well as secondary GBMs, a finding that offers new therapeutic approaches in these patients.
PMCID: PMC1907411  PMID: 17504928
low-grade glioma; methylation; PKB/Akt; PTEN; secondary glioblastoma
17.  Is the small heat shock protein αB-crystallin an oncogene? 
In the last 5 years, global gene expression profiling has allowed for the subclassification of the heterogeneous disease of breast cancer into new subgroups with prognostic significance. However, for most subgroups, the nature of the contributions of individual genes to the clinical phenotypes remains largely unknown. In this issue of the JCI, Moyano and colleagues further examine the oncogenic potential of the small heat shock protein α-basic–crystallin, commonly expressed in tumors of the basal-like breast cancer subtype associated with poor prognosis, and show that it is an oncogenic protein in the breast.
PMCID: PMC1323271  PMID: 16395401
18.  The New York cancer project: Rationale, organization, design, and baseline characteristics 
Cancer is the second leading cause of death in New York City, with nearly 15,000 deaths each year. The urban setting of New York City provides ready access to large and diverse populations for whom racial/ethnic disparities in cancer risk and outcomes can be examined. A new cohort study was undertaken with several aims: (1) to provide a database and biorepository for studies of cancer etiology and pathogenesis, including host genetics; (2) to differentiate risk factors that contribute to racial/ethnic disparities in cancer risk, prevention, control, incidence, mortality, and survival; (3) to provide timely data on cancer risk and preventive behaviors that can be used to mobilize and then evaluate public health programs. Scientists from multiple institutions contributed to protocol design and implementation. Study instruments included demographics, personal and family history of cancer, risk and prevention efforts. End points include linkage with registries and medical record reviews. Using venue-based sampling with quotas, 18,187 adults aged 30 years or older were recruited over a year to undergo a baseline questionnaire, venipuncture, and contact information. The sample was 39% male, 37% older than 50 years, 58% white, 20% African American, 18% Hispanic, and 9% Asian. In terms of family history of cancer, 21% reported mother, 21% reported father, and 5.9% reported both parents with cancer; 8.5% reported any sibling with cancer. At baseline, 1,231 participants reported prior cancer. Showing the feasibility of constructing a cohort based in New York City, plans proceed for additional recruitment and analyses on the salient questions about cancer.
PMCID: PMC3456454  PMID: 15136663
Cancer; Control; Epidemiology; Etiology; Incidence; Mortality; New York City; Pathogenesis; Prevention; Survival
19.  PTEN Expression Causes Feedback Upregulation of Insulin Receptor Substrate 2 
Molecular and Cellular Biology  2001;21(12):3947-3958.
PTEN is a tumor suppressor that antagonizes phosphatidylinositol-3 kinase (PI3K) by dephosphorylating the D3 position of phosphatidylinositol (3,4,5)-triphosphate (PtdIns-3,4,5-P3). Given the importance of PTEN in regulating PtdIns-3,4,5-P3 levels, we used Affymetrix GeneChip arrays to identify genes regulated by PTEN. PTEN expression rapidly reduced the activity of Akt, which was followed by a G1 arrest and eventually apoptosis. The gene encoding insulin receptor substrate 2 (IRS-2), a mediator of insulin signaling, was found to be the most induced gene at all time points. A PI3K-specific inhibitor, LY294002, also upregulated IRS-2, providing evidence that it was the suppression of the PI3K pathway that was responsible for the message upregulation. In addition, PTEN, LY294002, and rapamycin, an inhibitor of mammalian target of rapamycin, caused a reduction in the molecular weight of IRS-2 and an increase in the association of IRS-2 with PI3K. Apparently, PTEN inhibits a negative regulator of IRS-2 to upregulate the IRS-2–PI3K interaction. These studies suggest that PtdIns-3,4,5-P3 levels regulate the specific activity and amount of IRS-2 available for insulin signaling.
PMCID: PMC87057  PMID: 11359902
20.  Deficiency of Pten accelerates mammary oncogenesis in MMTV-Wnt-1 transgenic mice 
Germline mutations in the tumor suppressor PTEN predispose human beings to breast cancer, and genetic and epigenetic alterations of PTEN are also detected in sporadic human breast cancer. Germline Pten mutations in mice lead to the development of a variety of tumors, but mammary carcinomas are infrequently found, especially in mice under the age of six months.
To better understand the role of PTEN in breast tumor development, we have crossed Pten heterozygous mice to MMTV-Wnt-1 transgenic mice that routinely develop ductal carcinomas in the mammary gland. Female Wnt-1 transgenics heterozygous for Pten developed mammary tumors earlier than Wnt-1 transgenics that were wild type for Pten. In most tumors arising in Pten heterozygotes, the Pten wild-type allele was lost, suggesting that cells lacking Pten function have a growth advantage over cells retaining a wild type allele. Tumors with LOH contained high levels of activated AKT/PKB, a downstream target of the PTEN/PI3K pathway.
An animal model has been developed in which the absence of Pten collaborates with Wnt-1 to induce ductal carcinoma in the mammary gland. This animal model may be useful for testing therapies specific for tumors deregulated in the PTEN/PI3K/AKT pathway.
PMCID: PMC29091  PMID: 11178110

Results 1-20 (20)