Shinya Matsuzaki

Serum leucine-rich alpha-2 glycoprotein is a disease activity biomarker in ulcerative colitis

Background: Reliable biomarkers for monitoring disease activity have not been clinically established in ulcerative colitis (UC). This study aimed to investigate whether levels of serum leucine‐rich alpha‐2 glycoprotein (LRG), identified recently as a potential disease activity marker in Crohns disease and rheumatoid arthritis, correlate with disease activity in UC. Methods: Serum LRG concentrations were determined by enzyme‐linked immunosorbent assay (ELISA) in patients with UC and healthy controls (HC) and were evaluated for correlation with disease activity. Expression of LRG in inflamed colonic tissues from patients with UC was analyzed by western blotting and immunohistochemistry. Interleukin (IL)‐6‐independent induction of LRG was investigated using IL‐6‐deficient mice by lipopolysaccharide (LPS)‐mediated acute inflammation and dextran sodium sulfate (DSS)‐induced colitis. Results: Serum LRG concentrations were significantly elevated in active UC patients compared with patients in remission (P < 0.0001) and HC (P < 0.0001) and were correlated with disease activity in UC better than C‐reactive protein (CRP). Expression of LRG was increased in inflamed colonic tissues in UC. Tumor necrosis factor alpha (TNF‐&agr;), IL‐6, and IL‐22, serum levels of which were elevated in patients with active UC, could induce LRG expression in COLO205 cells. Serum LRG levels were increased in IL‐6‐deficient mice with LPS‐mediated acute inflammation and DSS‐induced colitis. Conclusions: Serum LRG concentrations correlate well with disease activity in UC. LRG induction is robust in inflamed colons and is likely to involve an IL‐6‐independent pathway. Serum LRG is thus a novel serum biomarker for monitoring disease activity in UC and is a promising surrogate for CRP. (Inflamm Bowel Dis 2012;)

Biomarkers for Screening, Diagnosis, and Monitoring of Ovarian Cancer

Serum tumor markers have a major role in the screening, diagnosis, and monitoring of most of the gynecologic cancers. Ovarian cancer is one of the deadliest of the group because it is so frequently asymptomatic until it has advanced to an untreatable stage. Even serum cancer antigen-125 (CA-125), clinically one of the most reliable serum markers for ovarian cancer, is elevated in only half of early-stage still-treatable tumors. Because of the very low prevalence of ovarian cancer in the general population, at present, there is no cost-effective imaging or simple microscopic screening test for ovarian cancer as there is for breast and cervical cancers. However, recent proteomics and nucleic acid–based analyses have shown great promise for the discovery of new and more useful serum biomarkers, which cumulatively might provide such a screening tool. In this review, we will discuss both the currently used serum tumor markers for screening, diagnosis, monitoring of ovarian cancer, and the novel biomarkers that are now under investigation and validation. Cancer Epidemiol Biomarkers Prev; 21(11); 1902–12. ©2012 AACR.

A Case of Disseminated Peritoneal Leiomyomatosis Developing after Laparoscope-Assisted Myomectomy

A 36-year-old nulliparous woman developed multiple extra-uterine fibroids in the pelvic cavity years after laparoscopic myomectomy. Molecular genetic analysis by methylation-specific polymerase chain reaction (MSPCR) of the human X-linked androgen receptor gene and loss of heterozygosity (LOH) analysis at 5 microsatellite loci was performed on the tumors. All tumors showed an identical non-random X-chromosome inactivation pattern by MSPCR and an identical pattern of LOH was found in all the tumors by LOH analysis. This demonstrated that 3 fibroids resected 2 years later and 14 fibroids resected 6 years later were all metastatic tumors originating from the uterine leiomyoma found during the initial surgery, suggesting that morcellation before removal of the leiomyoma nodule during laparoscopic myomectomy may have been associated with the pathogenesis of this case.

Expression of aldehyde dehydrogenase 1 (ALDH1) in endometrioid adenocarcinoma and its clinical implications

Aldehyde dehydrogenase 1 (ALDH1) is expressed in stem/progenitor cells, including cancer‐initiating cells (CIC) of various organs. In the present study, ALDH1 expression was immunohistochemically examined in uterine endometrioid adenocarcinoma. The ALDH1 was expressed in a small portion of tumor cells, and these ALDH1‐expressing cells were less mature than ALDH1‐non‐expressing cells. The ALDH1‐expressing (ALDH1‐hi) cells were more tumorigenic, resistant to anti‐cancer agents and more invasive than ALDH1‐lo cells. Culture of the sorted ALDH1‐hi cells yielded both ALDH1‐hi and ALDH1‐lo cells, whereas ALDH1‐lo cells yielded ALDH‐lo cells alone. Clinically, a high‐level of ALDH1 expression in tumor cells was correlated with T category, lymphatic invasion, recurrence and prognosis of patients. Patients with high ALDH1 expression showed poorer prognoses than those with low expression (P = 0.015 for disease‐free survival [DFS] and P = 0.010 for overall survival [OS]), and high ALDH1 expression was an independent factor for poor prognosis. Aldehyde dehydrogenase 1 is a candidate for CIC marker for uterine endometrioid adenocarcinoma. (Cancer Sci 2011; 102: 903–908)

Plasma membrane proteomics identifies bone marrow stromal antigen 2 as a potential therapeutic target in endometrial cancer

This report utilizes a novel proteomic method for discovering potential therapeutic targets in endometrial cancer. We used a biotinylation‐based approach for cell‐surface protein enrichment combined with isobaric tags for relative and absolute quantitation (iTRAQ) technology using nano liquid chromatography–tandem mass spectrometry analysis to identify specifically overexpressed proteins in endometrial cancer cells compared with normal endometrial cells. We identified a total of 272 proteins, including 11 plasma membrane proteins, whose expression increased more than twofold in at least four of seven endometrial cancer cell lines compared with a normal endometrial cell line. Overexpression of bone marrow stromal antigen 2 (BST2) was detected and the observation was supported by immunohistochemical analysis using clinical samples. The expression of BST2 was more characteristic of 118 endometrial cancer tissues compared with 59 normal endometrial tissues (p < 0.0001). The therapeutic effect of an anti‐BST2 antibody was studied both in vitro and in vivo. An anti‐BST2 monoclonal antibody showed in vitro cytotoxicity in BST2‐positive endometrial cancer cells via antibody‐dependent cell‐mediated cytotoxicity and complement‐dependent cytotoxicity. In an in vivo xenograft model, anti‐BST2 antibody treatment significantly inhibited tumor growth of BST2‐positive endometrial cancer cells in an NK cell‐dependent manner. The anti‐BST2 antibody had a potent antitumor effect against endometrial cancer both in vitro and in vivo, indicating a strong potential for clinical use of anti‐BST2 antibody for endometrial cancer treatment. The combination of biotinylation‐based enrichment of cell‐surface proteins and iTRAQ analysis should be a useful screening method for future discovery of potential therapeutic targets.

Intraoperative frozen section assessment of myometrial invasion and histology of endometrial cancer using the revised FIGO staging system.

Objectives: The objective of this study was to assess the value of intraoperative frozen section (IFS) diagnosis for myometrial invasion and histology of endometrial cancer using the revised International Federation of Gynecology and Obstetrics (FIGO) staging system. Methods: The medical records of 303 patients with endometrial cancer who underwent surgery with intraoperative diagnosis at the Osaka University Hospital between January 1999 and December 2008 were reviewed. Intraoperative frozen section diagnosis was retrospectively analyzed for the accuracy rates of myometrial invasion and histology compared with the final diagnosis and with preoperative prediction by magnetic resonance imaging (MRI) and endometrial curettage. Results: When using the previous FIGO staging system, the accuracy rate of IFS for the diagnosis of myometrial invasion was 77%, whereas the accuracy rate of preoperative prediction by MRI was 54%. However, using the newly revised FIGO staging system for myometrial invasion, the accuracy rate of IFS was 87% and the preoperative prediction by MRI was 82%. The accuracy rate of IFS for the diagnosis of histology was 71%, whereas the accuracy rate of preoperative prediction by endometrial curettage was 68%. Conclusion: Although under the previous FIGO staging system IFS diagnosis was significantly more accurate than preoperative prediction by MRI, when using the newly revised FIGO staging system, there are no significant differences between the values of preoperative and intraoperative diagnoses. The accuracy of IFS, however, trends to be slightly better than the preoperative procedures of MRI and endometrial surface biopsy. Thus, IFS diagnosis is still useful for directing primary operative management.

Annexin A4‐conferred platinum resistance is mediated by the copper transporter ATP7A

Although platinum drugs are often used for the chemotherapy of human cancers, platinum resistance is a major issue and may preclude their use in some cases. We recently reported that enhanced expression of Annexin A4 (Anx A4) increases chemoresistance to carboplatin through increased extracellular efflux of the drug. However, the precise mechanisms underlying that chemoresistance and the relationship of Anx A4 to platinum resistance in vivo remain unclear. In this report, the in vitro mechanism of platinum resistance induced by Anx A4 was investigated in endometrial carcinoma cells (HEC1 cells) with low expression of Anx A4. Forced expression of Anx A4 in HEC1 cells resulted in chemoresistance to platinum drugs. In addition, HEC1 control cells were compared with Anx A4‐overexpressing HEC1 cells in xenografted mice. Significantly greater chemoresistance to cisplatin was observed in vivo in Anx A4‐overexpressing xenografted mice. Immunofluorescence analysis revealed that exposure to platinum drugs induced relocation of Anx A4 from the cytoplasm to the cellular membrane, where it became colocalized with ATP7A, a copper transporter also well known as a mechanism of platinum efflux. ATP7A expression suppressed by small interfering RNA had no effect on HEC1 control cells in terms of chemosensitivity to platinum drugs. However, suppression of ATP7A in Anx A4‐overexpressing platinum‐resistant cells improved chemosensitivity to platinum drugs (but not to 5‐fluorouracil) to a level comparable to that of control cells. These results indicate that enhanced expression of Anx A4 confers platinum resistance by promoting efflux of platinum drugs via ATP7A.

Endometrial thickness measured by ultrasonography in postmenopausal patients with endometrial carcinoma has significance, irrespective of histological subtype.

Objective The criterion standard of practice for gynecologists is to measure the endometrial thickness with ultrasonography in women presenting with postmenopausal bleeding. A recent study reported that a thin endometrial stripe upon ultrasonography did not reliably exclude type II endometrial carcinoma. The aim of the present study was to reevaluate the reliability of ultrasonographic measurement of the endometrium for prediction of endometrial carcinomas of both types I and II in postmenopausal women. Methods We collected clinical data from patients with endometrial carcinoma who underwent surgical treatment at the Department of Obstetrics and Gynecology of the Osaka University Hospital, Osaka, Japan, during our study period from 2010 to 2012. Only the postmenopausal cases were included in our study. We excluded cases with insufficient clinical data. Results Preoperative measurement of the endometrium by transvaginal ultrasonography revealed that the endometrium was greater than 4 mm in 80 (89%) of the 90 type I cases and in 41 (93%) of the 44 type II cases. The median of the endometrial thickness measured with transvaginal ultrasonography preoperatively in type I cases, including both patients with myometrial invasion less than 1/2 and those with myometrial invasion greater than 1/2, was 13 mm (range, 1–78 mm). That of type II cases was 15 mm (range, 1–54 mm). This difference was not statistically significant (P = 0.46 by Mann-Whitney U test). These results implied that endometrial thickness was not significantly associated with the type of tumors. Conclusions Ultrasonographic measurements of the endometrium for prediction of endometrial carcinomas in postmenopausal women are reliable for both type I and type II tumors. These results encourage us to continue to use the “4-mm (5-mm) rule” to evaluate endometrial thickness in postmenopausal women, in opposition to a previous report.

Successful conservative management of placenta percreta: Investigation by serial magnetic resonance imaging of the clinical course and a literature review.

Placenta percreta is a rare condition that presents a risk of massive intraoperative hemorrhage. To decrease maternal morbidity in such cases, conservative management was recently adopted as an option. Nevertheless, severe morbidity occurs in 42% of cases. In addition, the clinical course of conservative management remains unclear. We encountered a case of placenta percreta that was successfully treated by conservative management without complications or other clinical interventions. Serial magnetic resonance imaging (MRI) was performed to investigate the natural course of conservative management of placenta percreta. MRI on postoperative day (POD) 99 showed a decrease in placental size without gadolinium enhancement. MRI on POD 162 showed that the residual placenta was approximately 4 cm in size without gadolinium enhancement. On POD 265, the placenta had almost disappeared. This is the first series of MRI of conservative management of placenta percreta without clinical interventions. We present our case and a literature review.

Anti-glypican-1 antibody-drug conjugate exhibits potent preclinical antitumor activity against glypican-1 positive uterine cervical cancer

Glypican‐1 (GPC1) is highly expressed in solid tumors, especially squamous cell carcinomas (SCCs), and is thought to be associated with disease progression. We explored the use of a GPC1‐targeted antibody‐drug conjugate (ADC) as a novel treatment for uterine cervical cancer. On immunohistochemical staining, high expression levels of GPC1 were detected in about 50% of uterine cervical cancer tissues and also in a tumor that had relapsed after chemoradiotherapy. Novel anti‐GPC1 monoclonal antibodies were developed, and clone 01a033 was selected as the best antibody for targeted delivery of the cytotoxic agent monomethyl auristatin F (MMAF) into GPC1‐positive cells. The anti‐GPC1 antibody was conjugated with MMAF. On flow cytometry, HeLa and ME180 cervical cancer cells highly expressed GPC1, however, RMG‐I ovarian clear cell cancer cell line showed weak expression. The GPC1‐ADC was rapidly internalized into GPC1‐expressing cells in vitro and was potently cytotoxic to cancer cells highly expressing GPC1. There were no inhibitory effects on cancer cells with low expression of GPC1. In a murine xenograft model, GPC1‐ADC also had significant and potent tumor growth inhibition. GPC1‐ADC–mediated G2/M phase cell cycle arrest was detected, indicating that the dominant antitumor effect in vivo was MMAF‐mediated. The toxicity of GPC‐ADC was tolerable within the therapeutic dose range in mice. Our data showed that GPC1‐ADC has potential as a promising therapy for uterine cervical cancer.