Tissue Counterparts Research Articles

Expression pattern of human SERPINE2 in a variety of human tumors.

Serine proteinase inhibitor, clade E member 2 (SERPINE2), also known as protease nexin-1 (PN-1), is a member of the serpin family. Despite several reported roles of SERPINE2 in tumor development the histological distribution of SERPINE2 and its expression levels in a large variety of tumors remains unclear. Through expressed sequence tag database analysis, immunohistochemical staining of tissue microarrays and a literature review, it was revealed that SERPINE2 expression varied according to growth stages and tissue types. SERPINE2 is differentially expressed in a number of tumors and their normal tissue counterparts. SERPINE2 is identified most abundantly in adenocarcinomas. SERPINE2 serves diverse roles in a variety of tumors and therefore may serve as a promising biomarker for tumor diagnosis and prognosis.

Characterization of Multilayered Tissue-Engineered Human Alveolar Bone and Gingival Mucosa

Advances in tissue engineering have permitted assembly of multilayered composite tissue constructs for potential applications in the treatment of combined hard and soft tissue defects and as an alternative in vitro test model to animal experimental systems. The aim of this study was to develop and characterize a novel three-dimensional combined human alveolar bone and gingival mucosal model based on primary cells isolated from the oral tissues. Bone component of the model was engineered by seeding primary human alveolar osteoblasts into a hydroxyapatite/tricalcium phosphate scaffold and culturing in a spinner bioreactor. The engineered bone was then laminated, using an adhesive tissue sealant, with tissue-engineered gingival mucosa consisting of air/liquid interface-cultured normal human gingival keratinocytes on oral fibroblast-populated collagen gel scaffold. Histological characterization revealed a structure consisting of established epithelial, connective tissue and bone layers closely comparable to normal oral tissue architecture. The mucosal component demonstrated a mature epithelium undergoing terminal differentiation similar to that characteristic of native buccal mucosa, as confirmed using cytokeratin 13 and cytokeratin 14 immunohistochemistry. Ultrastructural analysis confirmed the presence of desmosomes and hemidesmosomes in the epithelial layer, a continuous basement membrane, and newly synthesized collagen in the connective tissue layer. Quantitative polymerase chain reaction (qPCR) assessment of osteogenesis-related gene expression showed a higher expression of genes encoded collagen I (COL1) and osteonectin (ON) compared with osteocalcin (OC), osteopontin (OP), and alkaline phosphatase (ALP). Enzyme-linked immunosorbent assay quantification of COL1, ON, and OC confirmed a pattern of secretion, which paralleled the model's gene expression profile. We demonstrate in this study that, replicating the anatomical setting between oral mucosa and the underlying alveolar bone is feasible and the developed model showed characteristics similar to those of normal tissue counterparts. This trilayered model therefore offers great scope as an advanced and anatomically representative tissue-engineered alternative to animal models.

Overexpression of sprouty2 in human oral squamous cell carcinogenesis

ObjectiveThis study investigated SPRY2 expression in human oral potentially malignant disorders (OPMDs) and oral squamous cell carcinomas (OSCCs). Methods75 OSCCs, 23 OPMDs with malignant transformation (MT), 17 OPMDs without MT, and eight normal oral mucosa (NOM) tissues were used for immunohistochemical staining; three OSCC tissues with normal tissue counterparts were used for western blotting. Three human oral cancer cell lines (OCCLs), an oral precancer cell line (DOK), and a NOM primary culture (NOMPC) were used for western blotting; OCCLs and NOMPC were employed for real-time quantitative reverse transcription-polymerase chain reaction. OCCLs were evaluated in terms of proliferation, migration, invasion and BRAF V600E point mutation assays. ResultsSignificantly increased SPRY2 protein expression was observed in OSCCs as compared with NOM, and SPRY2 expression also differed between OSCC patients with and without lymph-node metastasis. SPRY2 protein and mRNA expressions were significantly enhanced as compared with NOMPC. Increased phospho-ERK expression was observed in OCCLs as compared with NOMPC. Significant decreases in the proliferation rate, degrees of migration and invasion were noted in OCCLs with SPRY2 siRNA transfection as compared with those without SPRY2 siRNA transfection. No BRAF V600E point mutation was observed for OCCLs as compared with NOMPC. A significantly increased SPRY2 protein level was noted in OPMDs with MT as compared to those without MT, and was also found in OPMDs with MT in comparison with NOM, as well as in DOK in comparison with NOMPC. ConclusionsOur results indicated that SPRY2 overexpression is associated with human oral squamous-cell carcinogenesis.

Enhanced inflammation and attenuated tumor suppressor pathways are associated with oncogene-induced lung tumors in aged mice.

SummaryAging is often accompanied by a dramatic increase in cancer susceptibility. To gain insights into how aging affects tumor susceptibility, we generated a conditional mouse model in which oncogenic Kras G12D was activated specifically in lungs of young (3–5 months) and old (19–24 months) mice. Activation of Kras G12D in old mice resulted in shorter survival and development of higher‐grade lung tumors. Six weeks after Kras G12D activation, old lung tissues contained higher numbers of adenomas than their young tissue counterparts. Lung tumors in old mice displayed higher proliferation rates, as well as attenuated DNA damage and p53 tumor suppressor responses. Gene expression comparison of lung tumors from young and old mice revealed upregulation of extracellular matrix‐related genes in young tumors, indicative of a robust cancer‐associated fibroblast response. In old tumors, numerous inflammation‐related genes such as Ccl7,IL‐1β, Cxcr6, and IL‐15ra were consistently upregulated. Increased numbers of immune cells were localized around the periphery of lung adenomas from old mice. Our experiments indicate that more aggressive lung tumor formation in older Kras G12D mice may be in part the result of subdued tumor suppressor and DNA damage responses, an enhanced inflammatory milieu, and a more accommodating tissue microenvironment.

GINGIVAL CYST OF ADULT WITH RADIOGRAFIC FINDINGS: A RARE CASE

Gingival cyst of adult (GCA) is an uncommon developmental cyst of gingiva arising from the rests of dental lamina. GCA occurs characteristically in the mandibular canine and premolar area and it is considered to represent the soft tissue counterpart of the lateral periodontal cyst (LPC). Radiologic changes occur in a small number of GCA. A 59-year-old female patient was referred to the University Hospital because of a firm and sessile nodule measuring 1.2 cm, located on the buccal papillae between the teeth 33–34. Radiographic analysis showed saucerization of the intercrestal bone with trabeculae within. The lesion was treated by surgical excision under local anesthesia. The histopathologic examination revealed a connective tissue capsule lined by thin non-keratinized squamous epithelium with focal nodular thickenings. The clinical diagnosis was established during surgical procedure, a depression in the outer cortical bone was observed in the LPC, the outer cortex is intact or expanded.

Decreased SIRT4 protein levels in endometrioid adenocarcinoma tissues are associated with advanced AJCC stage.

Members of the SIRT family are a highly conserved family of NAD+-dependent enzymes, many of which (SIRT1-7) play an important role in tumor formation. Recently, several studies have suggested that SIRT4 not only regulates glutamine metabolism, but also serves as a tumor suppressor. There are no studies have assessed its clinical significance in endometrioid adenocarcinoma. We investigated SIRT4 protein levels in endometrioid adenocarcinoma and its possible association with selected clinico-pathological parameters by immunohistochemical staining of a tissue microarray that included 65 endometrioid adenocarcinoma patients. SIRT4 protein levels in endometrioid adenocarcinoma were markedly lower than its non-neoplastic tissue counterpart (P< 0.001). Moreover, lower SIRT4 expression levels were observed in advanced AJCC stages of development (P= 0.002). Our results indicated that SIRT4 may be involved in the development of endometrioid adenocarcinoma and is a promising target for both the diagnosis and potential therapy of endometrioid adenocarcinoma.

Abstract 4338: The role of IMPACT in survival of cancer cells during tryptophan deprivation by immunosuppressive enzyme indoleamine 2,3-dioxygenase (IDO1)

Abstract A broad range of human malignancies overexpress an enzyme called indoleamine 2,3-dioxygenase 1 (IDO1) to suppress antitumor immunity which associates with poor patient prognosis. One mechanism whereby IDO1 activity suppresses the host’s immune cells is mediated by deprivation of the essential amino acid tryptophan (Trp) which raises an intriguing question. How do the cancer cells overcome low levels of the essential amino acid when the immune T-cells are triggered to self-destruct? A protein called IMPACT (product of IMPrinted gene with AnCienT domain) was reported to confer skin cells and neuronal cells increased survival in Trp-deprived environments. Based on these reports, we hypothesise that cancer cells hijack IMPACT to gain survival advantage in Trp-deprived environments. To obtain evidence for this hypothesis, we analysed publicly available genomics datasets and performed experimental assays. Murine GL261 glioblastoma cells overexpressing a full-length mouse IMPACT gene (GL261-IMPACT) were generated by lipofection. IMPACT overexpression was confirmed by Western blot. Metabolic activity and survival of the GL261 cells in Trp deprivation experiments were evaluated using MTT assay and staining with live/dead indicators fluorescein diacetate and propidium iodide, respectively. Meta-analysis of the publicly available RNA-sequencing datasets, The Cancer Genome Atlas (TCGA) and Genotype Tissue Expression (GTEx), provided evidence that IMPACT was overexpressed in the majority of the 24 analysed human cancers compared to their non-malignant tissue counterparts. Consistent with the high IMPACT expression in prostate cancer patients, 12-25% prostate tumors show amplification of the IMPACT gene (source: cBioPortal for Cancer Genomics). In addition, we found an association of high IMPACT expression with poorer survival for certain leukemia and lung cancer patients in the Gene Expression Omnibus (GEO) portal. To obtain experimental evidence that IMPACT protects cancer cells during Trp deprivation, GL261-IMPACT cells were cultured in media containing 2.5 - 50 μM Trp. Metabolic activity of GL261-IMPACT cells decreased at a slower rate than that of GL261-wild-type cells after 3 to 5 days in media containing limiting Trp concentration (2.5 - 10 μM). On day 5, GL261-wild-type and GL261-IMPACT cells cultivated in 10 μM Trp showed 4.5% and 23% metabolic activity (p = 2.8x10-8), respectively, relative to the same cells grown in 50 μM Trp. Fluorescence microscopy utilising live/dead staining showed that after 5 days of incubation at limiting Trp concentrations (5 - 10 μM), the majority of GL261-wild-type cells were non-viable whereas GL261-IMPACT cells were predominantly viable. Our initial results support the hypothesis that IMPACT aids cancer cells to overcome periods of tryptophan deprivation. Citation Format: Petr Tomek, Ariane Hallermayr, Michael Kilian, Cristin Gregor Print, Lai-Ming Ching. The role of IMPACT in survival of cancer cells during tryptophan deprivation by immunosuppressive enzyme indoleamine 2,3-dioxygenase (IDO1) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4338. doi:10.1158/1538-7445.AM2017-4338

Abstract 2733: Comparison of copy number aberrations (CNAs) between plasma cell free DNA (cfDNA) and tissue DNA in metastatic castrate resistant prostate cancer (mCRPC)

Abstract Background: Circulating cell-free DNA (cfDNA) analysis is emerging as a less invasive approach to assessing tumor genomic alterations in cancer patients. Although high concordance has been reported between tumor tissue NGS and cfDNA in studies investigating specific genetic alterations, the fidelity of cfDNA to tumor tissue DNA in the global genomic scale is largely unknown. In a correlative study of a prospective clinical trial (NCT# 01953640) conducted in mCRPC stage patients treated with abiraterone acetate/prednisone (A/P), we evaluated correlation of genomic CNA in tumor DNA obtained from biopsy of metastatic lesions and matched plasma cfDNA. Methods: mCRPC patients (pts) underwent two image-guided core biopsies of metastases prior to initiation of A/P (visit 1, V1) and 12 weeks after treatment (visit 2, V2). At both time points plasma was obtained at the time of the core biopsies and cfDNA was extracted using DNA Blood Mini Kit (Qiagen, Valencia, CA). High coverage (for tumor tissue) and low coverage (for cfDNA) whole genome sequencing reads were first mapped to the human genome hg19. Read counts (RC) from the mapped sequence files were then binned into 1Mb windows. The RC ratio in each genomic bin was calculated by comparing tumor tissue DNA to lymphocyte gDNA derived from the same patient, and was further log2 transformed, corrected for GC content, and normalized by CGHnormaliter. The fully normalized log2 ratios data was subjected to segmentation using DNAcopy algorithm. Results: Between 05/2013 and 09/2015, 92 patients (pts) were enrolled of which tissue and plasma NGS data both visits was available for 18 pts. The correlation of CNAs between tumor tissue and its cfDNA counterpart ranges from 0.013 to 0.83 for V1 samples, and -0.05 to 0.9 for V2 samples. The decreased correlation in some pairs of samples is largely due to low tumor content and heterogeneity in the cfDNA. Although there is a wide range of correlation, commonly shared CNAs were identified in multiple chromosomal regions, including loss in 8p, gain in 8q and chromosome 5 and X. On the other hand, several genomic regions show inconsistent CNAs between tumor tissue and cfDNA among 15 pairs of samples in which cfDNA have large tumor content. These include: loss in 6q, 1p, 2p, 9q, 11q, 13p, 15p, 20p and gain in 8q and 10p in cfDNA but not in tumor tissue; marked loss in chromosome 22 and gain in p arm of X chromosome in tumor tissue but much less evident in cfDNA. Conclusion: High concordance of CNAs between cfDNA and tumor tissue DNA can be achieved given sufficient tumor content of cfDNA. However, more CNAs can be identified in cfDNA than its tumor tissue counterpart. Our result suggests that cfDNA NGS is a useful tool to investigate clonal evolution associated with cancer progression. Citation Format: Chiang-Ching Huang, Meijun Du, Liguo Wang, Yen-chen Lin, Hua Huang, Liewei Wang, Liang Wang, Manish Kohli. Comparison of copy number aberrations (CNAs) between plasma cell free DNA (cfDNA) and tissue DNA in metastatic castrate resistant prostate cancer (mCRPC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2733. doi:10.1158/1538-7445.AM2017-2733

Abstract 3690: The leukocyte chemoattractant chemerin modulates PTEN via CMKLR1 in human tumors

Abstract Background: Recent data in preclinical models has shown that phosphatase and tensin homolog (PTEN) loss correlated with decreased tumor immune cell infiltration as well as decreased response to T cell-based immunotherapy. Chemerin (RARRES2) is a recently identified endogenous leukocyte chemoattractant shown to recruit innate immune cells through its G-protein coupled receptor CMKLR1. Chemerin/RARRES2 is commonly downregulated in prostate and other cancers (e.g. sarcoma) compared to their normal tissue counterparts. Methylome-wide studies in multiple tumor types have identified RARRES2 as being one of the most hypermethylated genes, potentially leading to decreased chemerin expression. Our previous preclinical studies showed that forced overexpression of chemerin in tumors was capable of recruiting immune effector cells, including T cells, into the tumor microenvironment and suppressing tumor growth. Methods: In order to study the effects of chemerin overexpression on tumor cell intrinsic processes we exposed prostate and sarcoma tumor lines to exogenous recombinant chemerin in vitro. Evaluation of PTEN was performed at both the mRNA and protein levels, using both quantitative PCR and Western blotting, in comparison with normal prostate epithelia RWPE-1 as well as PTEN-null PC3 cells as controls. in vitro invasion assays were performed to investigate the functional impact of chemerin exposure on tumor intrinsic activity. Knockdown of CMKLR1 using siRNA was performed to determine its role in tumor response. Results: Using both prostate and sarcoma tumor lines, we found exogenous chemerin was able to significantly upregulate PTEN expression at both the mRNA and protein levels in a dose-response manner. Exposure to chemerin did not result in increased apoptosis or altered in vitro proliferation. Importantly, chemerin treatment significantly decreased in vitro tumor invasion. Knockdown studies showed CMKLR1 abrogation resulted in restored tumor migration, suggesting a link between this GPCR and PTEN expression and activity. Conclusions: For the first time, to our knowledge, we have shown a link between chemerin and PTEN expression and activity in both prostate and sarcoma tumor lines. Our collective study shows chemerin’s ability to upregulate PTEN activity and to mitigate tumor cell migration via CMKLR1. This work has functional implications on both tumor cell intrinsic and extrinsic responses to chemerin-based immunotherapeutic strategies. Further studies are needed to investigate the interaction between chemerin and PTEN signaling, and its connection to oncogenic signaling pathways manipulated in malignant tumors. Additionally, future in vivo studies using these tumor lines will help further elucidate the chemerin-PTEN axis and its role in tumor immunosurveillance. We hypothesize that increased chemerin-driven PTEN activity may help facilitate an improved immunotherapy response and functional ability to attack cancer cells. Citation Format: Keith R. Rennier, Ping Wang, Robert Crowder, Russell K. Pachynski. The leukocyte chemoattractant chemerin modulates PTEN via CMKLR1 in human tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3690. doi:10.1158/1538-7445.AM2017-3690

Uterine Inflammatory Myofibroblastic Tumors Frequently Harbor ALK Fusions With IGFBP5 and THBS1.

Inflammatory myofibroblastic tumor (IMT) can occur in a number of anatomic sites, including the uterus. Like its soft tissue counterpart, uterine IMT frequently expresses ALK and harbors ALK genetic rearrangements. The aim of this study is to fully characterize the genetic fusions that occur in uterine IMT. We studied 11 uterine IMTs with typical histology and 8 uterine myxoid smooth muscle tumors (5 leiomyomas, 1 smooth muscle tumor of uncertain malignant potential, and 2 leiomyosarcomas) in which the differential of IMT was considered, using a RNA-sequencing-based fusion assay to detect genetic fusions involving ALK, ROS1, RET, NTRK1/3, and other genes. ALK was expressed in 10 of 11 IMTs and 1 tumor initially categorized as a myxoid leiomyoma (granular cytoplasmic staining with paranuclear accentuation). Fusion transcripts involving ALK were identified in 9 of 10 ALK immunopositive IMTs, with 3 harboring IGFBP5-ALK, 3 harboring THBS1-ALK, 2 harboring FN1-ALK, and 1 harboring TIMP3-ALK. Among the smooth muscle tumors, IGFBP5-ALK fusion transcript was identified in only 1 ALK immunopositive case. Further review revealed that although a diagnosis of IMT was considered for the ALK immunopositive myxoid leiomyoma, this diagnosis was not initially rendered only because fluorescence in situ hybridization analysis was interpreted as negative for ALK genetic rearrangement; this case is best reclassified as an IMT. Notably, all the ALK fusions identified in our study included the transmembrane domain-encoding exon 19 of ALK. Our findings confirm the high frequency of ALK fusions in uterine IMT, with an enrichment of novel 5' ALK fusion partners (IGFBP5, THBS1, and TIMP3) and exon 19-containing ALK fusion. Given that IGFBP5 and FN1 are both situated on the same chromosome as ALK, fluorescence in situ hybridization analysis for ALK rearrangement may not be reliable and a negative result should not exclude a diagnosis of uterine IMT if the histologic features and ALK immunostaining findings are supportive.

Pericytic tumors of the kidney—a clinicopathologic analysis of 17 cases

The pericytic (perivascular myoid cell) family of tumors is a distinctive group of mesenchymal neoplasms encountered in superficial sites and only rarely seen in viscera. The pericytic family subtends a spectrum of lesions, namely, glomus tumors and variants; myopericytoma, including myofibroma; and angioleiomyoma. In light of the contemporary classification of pericytic lesions, we identified and reviewed 17 cases of renal pericytic tumors from the files of 6 referral centers. These tumors presented over an age range of 17 to 76 years (mean 46.7, median 53), with essentially equal male-female ratio. History of hypertension (available in 11 patients) was noted in 7 (64%), which persisted even after surgical resection, including in 2 younger patients (17 and 30 years). The tumors (1.7-11.0 cm) included glomus tumors (n=11); glomangiomyoma (n=1); glomus tumor with atypical features (n=1); and angioleiomyoma (n=1), as well as tumors showing features overlapping pericytic tumor subtypes (n=3). The histomorphology observed in these renal examples closely resembled that of their soft tissue counterparts, a subset with symplastic changes and atypical features, and pericytic immunophenotype. Despite large size and deep site, no progression was identified during a median of 7 months follow-up (1-62 months). In context of prior reported experience, our series identifies a wide morphologic spectrum, including lesions presenting composite morphologies. Taken with the experience of others, our series further corroborates that malignant behavior is rare, and that criteria associated with aggression among soft tissue pericytic tumors may not be predictive for those in the kidney.

Primary Ewing Sarcoma / Primitive Neuroectodermal Tumor of the Kidney: A Clinicopathologic Study of 23 Cases.

Primary Ewing sarcoma / primitive neuroectodermal tumor (ES) of the kidney is a rare neoplasm with limited clinicopathologic data. We report 23 such cases with no history of ES elsewhere in the body. The patients included 13 male and 10 female, aged 8-70years (mean, 31years). The average tumor size was 11.7cm (range, 5-20cm). Microscopic analysis showed predominantly lobular growth (n=14), with focal papillary (n=3), alveolar (n=1), and hemangiopericytoma-like (n=1) patterns. Several tumors (n=11) exhibited robust mitotic activity (>10 mitoses/10 high-power fields). Necrosis (n=13) and lymphovascular invasion (n=14) were common. Homer Wright rosettes (n=6) and perivascular pseudorosettes (n=1) were also identified. The tumors invaded the renal sinus or perinephric fat (n=11), renal vein (n=13), and adrenal gland (n=2). Molecular and fluorescence in situ hybridization analysis showed rearrangement of EWSR1 gene (10/10), associated with EWSR1-FLI1 gene fusion (7/10). All patients with follow-up information (n=18) had metastasis, commonly in the lungs (n=12) and bone (n=6). Twelve patients died of disease in a mean of 21months; 6 patients were alive at a mean of 49months after diagnosis. Primary kidney ES usually present at an advanced stage with extrarenal spread and metastasis. Although renal ES share histologic, immunohistochemical, and molecular features with their bone and soft tissue counterparts, they appear to be more aggressive tumors with poorer clinical outcome.

Oncopression: gene expression compendium for cancer with matched normal tissues

Expression profile of normal tissue is primary source to find genes showing aberrant expression pattern specific in matched cancer tissue, but sample number of normal control in public gene expression repositories is disproportionally small compared to cancer and scattered in several datasets. We built oncopression by integrating several datasets into one large dataset for comprehensive analysis about 25 types of human cancers including 20 640 cancer samples and 6801 normal control profiles. Expression profiles in cancers can be directly compared to normal tissue counterparts. Validity of the integration was tested using immunohistochemical staining results and principal component analysis. We have utilized the pre-release version of oncopression to identify cancer-specific genes in several studies. Free access at http://www.oncopression.com and all expression data are available for download at the site. cchoi@kaist.ac.kr or jungsullee@gmail.com. Supplementary data are available at Bioinformatics online.

Abstract P1-02-12: Determination of a serum progranulin (GP88/PGRN) level associated with overall survival in metastatic breast cancer patients

Abstract Imaging technologies are the methods of choice in the standard of care (SOC) to monitor therapy response in metastatic breast cancer (MBC) patients. However, such methods are expensive and have limited sensitivity to detect disease response in a timely manner. Measurement of the circulating tumor markers such as CA15-3, CA27.29 and CEA has provided additional minimally invasive methods in disease management of MBC patients. While useful, they have limitations in providing clinicians with a reliable insight into real-time disease monitoring. Understanding of real-time biological processes may provide better biomarkers of the disease state and thus aid real-time clinical management of MBC patients by identifying circulating disease associated biomarkers. Thus, addition of such new circulating biomarkers may improve the management of MBC patients. We have characterized a target biomarker that would fit these criteria, the 88kDa glycoprotein Progranulin (GP88). GP88 is expressed in tumor tissue and not in normal mammary tissue counterpart and is secreted in the circulation of BC patients. Biological studies have established GP88 as one of the critical drivers for breast cancer cell proliferation, survival, invasiveness and drug resistance. Clinical studies have demonstrated that elevated GP88 tumor levels were prognostic for recurrence and that breast cancer patients had a statistically elevated GP88 serum level than healthy individuals. Using tissue and serum tests to detect and quantify GP88 could provide an new strategies for identifying patients at high risk of recurrence and monitoring disease progression in BC patients undergoing therapy. In the present study, we examined whether GP88 serum levels were elevated in MBC patients and whether GP88 serum levels were correlated to patient overall survival. Under an IRB approved protocol, 92 MBC patients that met the inclusion criteria and were undergoing therapy at the UMGCCC Breast Clinic were consented and enrolled. Clinical and disease characteristics along with serum CA15-3 values were collected as part of the study. Serum samples were collected from each patient during therapy and subsequently the patients were monitored. The serum was stored at -80C until tested for GP88 using a GP88 enzyme linked immunoassay developed in our laboratory. Statistical analysis using Kaplan-Meier functions established whether there was a correlation between GP88 serum level and overall survival in MBC patients. By analyzing the KM plots at different GP88 cut points, we identified two populations with distinct survival characteristics. When examined more thoroughly the difference in overall survival of patients with &amp;lt;60ng/ml and &amp;gt;60ng/ml was statistically significant (P=0.0002). Correlation analysis of serum GP88 and CA15-3 were performed and will be presented. We conclude that circulating levels of GP88 in MBC patients are correlated with overall survival. It would appear that patients that can be managed to have a GP88 below 60ng/ml will survive longer. Thus measuring circulating GP88 levels would provide additional information to that available in today's SOC for monitoring. This valuable insight into real-time disease status will assist clinicians in patient management. Citation Format: Serrero G, Hawkins DM, Yue B, Hicks D, Tait N, Tkaczuk KR. Determination of a serum progranulin (GP88/PGRN) level associated with overall survival in metastatic breast cancer patients [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-02-12.

Primary extraskeletal myxoid chondrosarcoma of bone: Report of three cases and review of the literature

Extraskeletal myxoid chondrosarcoma is a rare neoplasm of soft tissue. The usual location is in deep parts of the proximal extremities and limb girdles in middle-aged adults. The bone location as primary location is extremely rare and few cases are reported. We present three cases arising in bone with molecular confirmation using both RT-PCR and FISH analysis. Patients include two men and one woman with an age of 62, 69 and 73 years old. The mean size of the lesion was 13cm (range 8–18cm). Tumors arose in the iliac bone in two cases and in the proximal humerus in the other case. At time of diagnosis the three cases show bone cortex and soft tissue involvement. On imaging, lesions have a lobular pattern, are purely lytic, but take up contrast medium after injection. Two patients are alive with disease (local recurrence and lung metastasis) after five years and five years and six months, respectively and one patient died of disease two years after the diagnosis. The primary extraskeletal myxoid chondrosarcoma of bone seems to have a more aggressive behavior than the soft tissue counterpart. The molecular confirmation of diagnosis using RT-PCR is necessary to do the differential diagnosis with other entities, in particular with myoepithelioma that shows similar morphological features and EWSR1 and FUS genes rearrangement.

Oncogenic role of rab escort protein 1 through EGFR and STAT3 pathway

Rab escort protein-1 (REP1) is linked to choroideremia (CHM), an X-linked degenerative disorder caused by mutations of the gene encoding REP1 (CHM). REP1 mutant zebrafish showed excessive cell death throughout the body, including the eyes, indicating that REP1 is critical for cell survival, a hallmark of cancer. In the present study, we found that REP1 is overexpressed in human tumor tissues from cervical, lung, and colorectal cancer patients, whereas it is expressed at relatively low levels in the normal tissue counterparts. REP1 expression was also elevated in A549 lung cancer cells and HT-29 colon cancer cells compared with BEAS-2B normal lung and CCD-18Co normal colon epithelial cells, respectively. Interestingly, short interfering RNA (siRNA)-mediated REP1 knockdown-induced growth inhibition of cancer cell lines via downregulation of EGFR and inactivation of STAT3, but had a negligible effect on normal cell lines. Moreover, overexpression of REP1 in BEAS-2B cells enhanced cell growth and anchorage-independent colony formation with little increase in EGFR level and STAT3 activation. Furthermore, REP1 knockdown effectively reduced tumor growth in a mouse xenograft model via EGFR downregulation and STAT3 inactivation in vivo. These data suggest that REP1 plays an oncogenic role, driving tumorigenicity via EGFR and STAT3 signaling, and is a potential therapeutic target to control cancers.

Long non‑coding RNA AB019562 promotes cell proliferation and metastasis in human hepatocellular carcinoma.

Increasing evidence has supported the prognostic and therapeutic values of long non-coding RNAs (LncRNAs) in human tumorigenesis. Hepatocellular carcinoma (HCC), as one of the most refractory diseases, continues to warrant investigation for novel clues to enable early diagnosis. In the present study, the role of LncRNA AB019562 in cell proliferation and metastasis was investigated in HCC. Reverse transcription-quantitative polymerase chain reaction analysis was performed to determine the expression of AB019562 in clinical HCC samples and cultured HCC cells. In addition, a specific small interfering RNA against AB019562 was designed and transfected into HCC cells. A3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and a transwell assay were used to assess the effects of AB019562 knockdown on cell proliferation and metastasis, respectively. The results revealed that the expression of AB019562 was increased 4-fold in the clinical HCC tissues, compared with adjacent non-cancerous tissue counterparts. AB019562 was differentially expressed in the HCC cell lines. The knockdown of AB019562 reduced the rate of cell proliferation by 28.6% in HepG2 cells and by 24% in SMMC-7721 cells. Cell cycle assays revealed that the proportion of cells in the G0/G1 phase was significantly increased, whereas those in the S and G2/M phases were decreased in the AB019562-knockdowncells. The results of the transwell assay showed that the knockdown of AB019562 inhibited cell migration abilities by up to 67% in the HepG2 cells and 63% in the SMMC-7721 cells, and significantly suppressed invasive abilities by up to 75% in the HepG2 cells and 60% in the SMMC-7721 cells. Furthermore, AB019562 knockdown increased the apoptotic rates of the two cell lines and activated the expression of caspase-3, but not caspase-8. These data demonstrated the pro-oncogenic properties of AB019562 and suggested that AB019562 may serve as a novel biomarker for the diagnosis and treatment of patients with HCC.

Imaging review of lipomatous musculoskeletal lesions.

Lipomatous lesions are common musculoskeletal lesions that can arise within the soft tissues, bone, neurovascular structures, and synovium. The majority of these lesions are benign, and many of the benign lesions can be diagnosed by radiologic evaluation. However, radiologic differences between benign and malignant lipomatous lesions may be subtle and pathologic correlation is often needed. The use of sonography, computed tomography (CT), and magnetic resonance imaging (MRI) is useful not only in portraying fat within the lesion, but also for evaluating the presence and extent of soft tissue components. Lipomas make up most soft tissue lipomatous lesions, but careful evaluation must be performed to distinguish these lesions from a low-grade liposarcoma. In addition to the imaging appearance, the location of the lesion and the patient demographics can be utilized to help diagnose other soft tissue lipomatous lesions, such as elastofibroma dorsi, angiolipoma, lipoblastoma, and hibernoma. Osseous lipomatous lesions such as a parosteal lipoma and intraosseous lipoma occur less commonly as their soft tissue counterpart, but are also benign. Neurovascular and synovial lipomatous lesions are much rarer lesions but demonstrate more classic radiologic findings, particularly on MRI. A review of the clinical, radiologic, and pathologic characteristics of these lesions is presented.

Free radical scavenging injectable hydrogels for regenerative therapy

Pathological free radicals generated from inflamed and infarcted cardiac tissues interferes natural tissue repair mechanisms. Hypoxic microenvironment at the injured zone of non-regenerating cardiac tissues hinders the therapeutic attempts including cell therapy. Here we report an injectable, cytocompatible, free radical scavenging synthetic hydrogel formulation for regenerative therapy. New hydrogel (PEAX-P) is prepared with D-xylitol-co-fumarate-co-poly ethylene adipate-co-PEG comaromer (PEAX) and PEGDiacrylate. PEAX-P hydrogel swells 4.9 times the initial weight and retains 100.07kPa Young modulus at equilibrium swelling, which is suitable for cardiac applications. PEAX-P hydrogel retains elastic nature even at 60% compressive strain, which is favorable to fit with the dynamic and elastic natural tissue counterparts. PEAX-P hydrogel scavenges 51% DPPH radical, 40% hydroxyl radicals 41% nitrate radicals with 31% reducing power. The presence of hydrogel protects 62% cardiomyoblast cells treated with stress inducing media at LD 50 concentration. The free hydroxyl groups in sugar alcohols of the comacromer influence the free radical scavenging. Comparatively, PEAX-P hydrogel based on xylitol evinces slightly lower scavenging characteristics than with previously reported PEAM-P hydrogel containing mannitol having more hydroxyl groups. The possible free radical scavenging mechanism of the present hydrogel relies on the free π electrons associated with uncrosslinked fumarate bonds, hydrogen atoms associated with sugar alcohols/PEG and radical dilution by free water in the matrix. Briefly, the present PEAX-P hydrogel is a potential injectable system for combined antioxidant and regenerative therapy.

Higher expression of XPF is a critical factor in intrinsic chemotherapy resistance of human renal cell carcinoma.

Human renal cancer is extremely resistant to chemotherapy and radiation therapy. This clinical characteristic reduces the efficacy of chemotherapeutic agents in the treatment of recurrence or metastasis following surgical resection. Understanding the mechanism of chemotherapy resistance in renal cell carcinoma remains a significant challenge. In this study, we have shown that varied level of XPF expression was organ-tissue specific by comparing human renal cancer, bladder cancer, testicular cancer and their normal tissue counterparts, respectively. The expression of XPF was significantly higher in renal cancer than in bladder cancer and testicular cancer and correlated with the clinical characteristic of their chemotherapeutics sensitivity. These novel findings proposed that the intrinsic chemoresistance of human renal cell carcinomas might be derived from the high level of XPF expression. In a panel of five cancer cell lines, decreasing cisplatin sensitivity correlated with increasing levels of XPF expression. Knockdown of XPF expression not only increased sensitivity of renal carcinoma cells to cisplatin treatment by affecting the DNA damage response, including DNA repair, cell cycle regulation and apoptosis, but also increased senescence of renal cancer cell. Furthermore, experiment in vivo confirmed that silenced XPF significantly increased the sensitivity and survival following treatment with cisplatin in xenograft mice bearing renal cell tumor. These findings firstly uncover a partial mechanism of intrinsic chemoresistance in renal cancer and may provide a new approach to break through the obstacle of intrinsic chemoresistance by targeting the XPF protein with a potential new inhibitor.