Multi-omic biomarker detection in ovarian cancer.

Automated quantitative analysis of DCC tumor suppressor protein in ovarian cancer tissue microarray shows association with β-catenin levels and outcome in patients with epithelial ovarian cancer

Ovarian cancer typically presents late and has a poor prognosis. Ovarian screening aims to reduce mortality from the disease by detecting and treating women at an earlier, presymptomatic stage. The paper by Menon et al.1 published in BJOG more than a decade ago, set the stage for a multimodal approach to ovarian cancer screening, whereby a combination of CA125 levels and ultrasound scan findings could identify women at risk of the disease. Subsequent studies have not only informed the current management of women with suspected ovarian cancer but will also determine whether ovarian cancer screening can save lives. This commentary documents the ovarian screening story so far from inception to current day as we await the results from UKCTOCS, the largest ovarian cancer screening trial conducted to date. Ovarian cancer is known as the ‘silent killer’ because it only becomes symptomatic in its late stages. Compared with other common cancers, women diagnosed with ovarian cancer have a relatively poor prognosis. The overall 5-year survival rate of 42.9% masks vast differences in prognosis for those women presenting with Stage I disease compared with those with Stage IV disease.2 Since the late 1980s there has been a steady improvement in 5-year survival in the relatively small group of women who present with Stage I disease from 80% in 1987–91 to 92% in 2005–08.3 Changes in survival for the much larger proportion of women who present with Stage III and IV disease have been more modest. The 5-year survival rate for those with stage IV disease, however, stands unchanged at around 5%.3 These dismal survival rates for most women with ovarian cancer have stimulated enormous research effort aimed at identifying and treating women with early-stage disease. The hypothesis is that ovarian cancer screening may pick up early, asymptomatic disease in apparently healthy women that is then more amenable to surgical cure than disease that presents with symptoms. A major challenge when screening an asymptomatic population is that the proportion of true positives is extremely low given that the age-standardised incidence of ovarian cancer is only 16 per 100 000.4 Furthermore, the consequence of a positive screen is referral for laparoscopy/laparotomy with its incumbent risks. Hence, it has been suggested to ensure an acceptable false-positive rate any test needs to achieve a specificity of over 99.6%.5 This equates to nine women being offered further investigation or treatment for every woman diagnosed with an ovarian malignancy. This degree of specificity has not been achieved by any individual marker alone. To overcome this obstacle a number of strategies incorporating multiple markers have been proposed since the 1980s. The CA125 antigen was discovered in the early 1980s as part of an effort to develop a monoclonal antibody that could be used for immunotherapy of ovarian cancer.6 In this regard CA125 appeared to be relatively specific to epithelial ovarian cancers, being expressed in up to 80% of these cancers. Furthermore, it was found in the serum of women with ovarian cancer, although it can also be raised in a number of other cancers as well as some benign conditions. Although targeting CA125 has not proved to be therapeutically successful,7, 8 it was to prove a much more useful diagnostic/predictive biomarker. Its promise as a marker of early disease was shown in studies led by Zurawski in the 1980s. Serum CA125 levels were rarely elevated in healthy postmenopausal women9 but in up to 20% of cases, raised CA125 could be seen at least 2 years before ovarian cancer diagnosis.10 Alone, a raised CA125 has a high specificity but the likelihood of a woman having an occult cancer with a raised CA125 (Positive Predictive Value; PPV) remains low. PPV and sensitivity are determined in large part by the cut-offs used. Early studies by Klug et al. had determined that if a cut-off of 35 U/ml was used, then 82% of women with cancer, 6% of women with benign disease and 1% of normal women would have a raised CA125. Increasing the cut-off to 65 U/ml saw the percentage of ovarian cancer patients with a positive CA125 fall to 73% but those with benign disease and normal disease drop to 2% and 1%, respectively. To ensure that no women with benign disease would be included would necessitate the reference value to be set at 220 U/ml, which would mean that nearly 40% of women with ovarian cancers would be missed. Clearly serum CA125 measurement as a sole test was unlikely to offer the assurance of a low false-positive and false-negative rate needed to roll-out population screening. The 1950s saw the development of medical ultrasound and with it the ability to characterise gynaecological masses. Fittingly it was one of the first uses demonstrated by the seminal work of Ian Donald, Professor of Midwifery and a pioneer of diagnostic ultrasound.11 However, it would not be until the 1970s that this diagnostic tool was perceived to be a viable method for early detection in its own right, rather than a simple adjunct to the clinical examination of the symptomatic woman. Advances in ultrasound technology meant that by the mid-1970s lesions of 1 cm in diameter, which would normally be undetectable clinically, could be defined by ultrasound.12 Furthermore, various criteria to distinguish benign from malignant lesions with accuracies ranging from 70 to 90% were proposed.13-15 In 1982, Campbell et al.16 suggested that because ultrasound was able to define ovarian size accurately, ovarian volume could be a useful discriminator for potential ovarian disease in postmenopausal women. This strategy was studied in a large-scale prospective study of 5479 women followed over 8 years.17 Campbell et al. were able to identify 100% of the women that developed primary ovarian cancer within this time frame. The specificity of this procedure was 97.1% with a PPV of 1.5%, i.e. odds of a positive screen result representing a primary cancer of 1 in 67. Although Campbell et al. were able to demonstrate the utility of transabdominal ultrasound to identify women with asymptomatic ovarian masses, the inability to satisfactorily distinguish those with benign from malignant lesions underlies the high false-positive rates. The first approach proposed to reduce false positives was that only women with increasing ovarian volumes at a second scan, who had previously demonstrated either ovarian volumes >96th centile or abnormal morphology, e.g. cystic areas, should be offered surgery.18 Under this strategy, the PPV rose to 2.0%. The second approach came with the advent of transvaginal and Doppler ultrasound. Transvaginal ultrasound provided a higher resolution and so had the ability to detect abnormal morphology in smaller ovaries, while Doppler ultrasound provided the ability to visualise areas of neovascularisation more common in malignant lesions. Small studies showed promising results for the detection of women with ovarian pathology and the distinction of benign from malignant disease using transvaginal ultrasonography.19 The University of Kentucky Ovarian Cancer Screening Project was the first large-scale prospective trial to investigate whether transvaginal ultrasound assessment of ovarian volume could be used in ovarian cancer screening in pre- and postmenopausal women. In total, 90 women (1.4%) with persistent abnormalities were offered surgery to remove the affected ovary; six of whom were discovered to have a primary ovarian cancer. Overall, this strategy resulted in a sensitivity of 85.7%, specificity of 98.7% and 14 false positives for every woman with screen-detected ovarian cancer. Forgoing the possibility that the removal of benign ovarian masses with the potential for malignant transformation may lead to a resultant late fall in mortality from ovarian cancer, there remained a need for strategies to reduce the proportion of false positives. Jacobs et al.5 were the first to describe a multimodal approach for ovarian cancer screening in 1988. In their initial study, 1010 healthy postmenopausal women were offered serum CA125 measurement and a clinical pelvic examination. If either the clinical or biochemical assessment was abnormal women were offered a transabdominal ultrasound scan to assess ovarian volume. When CA125 measurements were considered individually, specificity was 97%, but when ultrasonography was added as a secondary test, specificity increased to 99.8%. This two-stage strategy was validated by a larger prevalence study in which 22 000 women were screened. With this approach equally high specificity and PPVs were achieved for a sensitivity of 79% at 1 year of follow up when a cut-off of 30 U/ml was used.20 The same cohort was then randomised to annual incidence screening or no further screening and followed up over 8 years. Initially, women recruited to this study in the late 1980s were offered transabdominal ultrasonography on entry. Subsequently, as evidence of the superiority of the transvaginal route in terms of image resolution with high acceptability among patients became available, follow-up scans were offered transvaginally. In contrast to the Kentucky Ovarian Cancer Screening Project, only postmenopausal women who had an abnormal CA125 were offered an ultrasound. In this preselected group of women, ultrasonographical assessment led to a much higher PPV of 21.3% and 23.75%, respectively, when ovarian volume and morphology were considered separately.1 Furthermore, there appeared to be minimal loss in sensitivity or specificity with this approach (sensitivity 89.5% and 100%; specificity 93.75% and 93.95% when considering ovarian volume and morphology, respectively). This was a breakthrough as it demonstrated that the target of nine healthy women being investigated or treated for every woman with cancer identified by screening was not only attainable but could be improved upon. This landmark finding stimulated a new wave of research to refine the detection of early ovarian cancers by screening. While early detection is fundamental, it is only relevant if it can translate into lives saved. Further analysis showed that screening identified proportionately more early-stage ovarian cancer (31.3% versus 10%), although the difference between the two groups was not statistically significant (P = 0.171).21 Nevertheless, the histological grade was lower and length of survival following randomisation was longer for women with ovarian cancer in the screened group (P = 0.02 and P = 0.01, respectively). Despite large numbers, the study was underpowered to detect a difference in mortality among screened and unscreened women, setting the scene for a still larger trial to answer this crucial question. Until 1998 there had been 25 prospective studies of ovarian cancer screening but no comparison with an unscreened population.22 Both ultrasound-based and multimodal screening strategies appeared to be able to identify asymptomatic women with ovarian cancer at early stages, but the clinical significance of this in terms of survival and mortality remained unknown. Furthermore ultrasound-based techniques appeared to offer higher sensitivity although at the cost of lower specificity, than multimodal screening. Notably, most of the women who received diagnostic surgery as a result, were found to have a benign gynaecological condition. Again the clinical impact of surgery in these circumstances is unclear. To answer this question UKCTOCS (UK Collaborative Trial of Ovarian Cancer Screening), the largest randomised control trial to date of an ovarian cancer screening strategy, involving over 200 000 women followed over 7 years was established. This study primarily aimed to ascertain whether screening, be it by a multimodal or imaging-only approach, could have an impact on ovarian cancer mortality. The trial would also assess the physical and psychological impact of screening, volunteer compliance and cost-effectiveness to inform decisions about the use of a national screening programme. As a result of further analysis of data from the pilot study, the multimodal arm had a new weapon in its armamentarium: the Risk of Ovarian Cancer algorithm (ROCA).23 This algorithm stratifies patients according to their CA125 trajectory rather than using a single threshold of 30 U/ml and is recalculated with every successive CA125 result. It was designed to improve sensitivity for the detection of early-stage cancers where earlier intervention could offer the most survival benefit and where CA125 is >30 U/ml in only 57% of cases, compared with 84% of late-stage cancers.24 Using the ROCA, a woman with a low baseline CA125 that suddenly rose would be identified as at risk of ovarian cancer, even if the absolute level remained modest, while a woman with a static but high CA125 would be deemed low risk. Retrospective testing of the algorithm on a Swedish cohort demonstrated high levels of sensitivity and specificity, which were subsequently verified by a prospective cohort of over 16 000 women between 1995 and 2000.25-27 More recently, ROCA was employed by Lu et al.28 in a large single-arm prospective study, which like UKCTOCS triaged women deemed high risk by ROCA to transvaginal ultrasound. Encouragingly, this protocol resulted in <1% of women being referred for surgery, giving it a specificity of 99.9% (95% confidence interval 99.7–100%) and a PPV of 40% (95% confidence interval of 12.2–78%).28 This equates to just over two women requiring surgery for every woman diagnosed with cancer. The pilot study published in BJOG,1 although primarily aimed at providing the platform for general population screening for postmenopausal women in whom 90% of ovarian cancer occurs, also provided the basis for studies in a second group of women with a family history of ovarian cancer. Some women are diagnosed with gene faults that increase their risk of ovarian and also breast cancer, notably defects in the BRCA1 and BRCA2 genes, but others have unidentified gene faults and/or are labelled as ‘at risk’ by virtue of their strong family history. Given the poor prognosis of ovarian cancer presenting symptomatically, women at high risk of ovarian cancer are currently offered risk-reducing prophylactic salpingo-oophorectomy (RRSO) to prevent incident ovarian cancers and reduce their risk of premenopausal breast cancer. When performed in young women, such surgery inevitably results in infertility and premature menopause and so screening may facilitate women safely delaying surgery until after their natural menopause. The United Kingdom Familial Ovarian Cancer Screening Study (UKFOCSS) recruited 3563 women with a 10% or greater lifetime risk of ovarian cancer between 2002 and 2008 to determine whether screening could be a viable alternative to RRSO. Reports from the first phase of the UKFOCSS study suggest that a negative screen can reliably reassure women (99.9% probability) that they will not develop ovarian cancer in the subsequent year. Screening was able to detect incident disease with a sensitivity of >80% and may reduce the proportion of stage IIIc disease.29 Phase II of the study drew on the successes of the UKCTOCS by stratifying patients according to ROCA using a computerised system, which would prompt investigators to repeat blood tests in women with intermediate risk and organise scans/referrals in women who were high risk. Additionally, screening intervals were reduced to 4 months to reduce false negatives.29 These amendments to the study design resulted in no interval cancers in this phase with no significant increase in the proportion of women referred.30 PPV in this phase was 13%. All women diagnosed with ovarian cancer as a result of screening went on to have optimal cytoreduction at surgery and survival was increased by 43 months.30 However, until such time as the efficacy of screening in this context has been fully evaluated, RRSO will remain the recommended intervention in women with a genetic predisposition to ovarian cancer. This research1 has also benefitted women with suspected ovarian cancer. The search for screening strategies which could be translated from women with clinically apparent cysts to those with occult disease fortuitously led to the development of the ‘Risk of Malignancy’ Index (RMI).31 This simple algorithm combines three factors that are independently predictive for an increased risk of ovarian cancer; menopausal status (scored 1or 3), serum CA125 and ultrasound features of the cyst (scored 0, 1 or 3). The product of these three criteria results in a score that can be used to provide a quantitative and reproducible assessment of likelihood that a symptomatic patient does indeed have an ovarian malignancy. Cut-offs can be useful tools for clinical decision making, e.g. referral to tertiary gynaecological oncology centres for surgery. To put this into context an RMI of 200 equates to a likelihood ratio of 42.1 of malignant disease and a likelihood ratio of 0.22 of benign disease; a 280-fold difference.32 Although the effect of the use of the RMI on the prognosis of a woman diagnosed with ovarian cancer has not been formally assessed, many studies have demonstrated the survival benefit of being treated by a specialist gynaecological oncologist.33 Where resources are limited, adjusting cut-offs can ensure only those at highest risk will be referred to tertiary centres. RMI remains the most widely used and evidenced stratification tool34 and in the UK, National Institute for Health and Care Excellence (NICE) guidance suggests the use of this tool to assess all women presenting to primary and secondary care with suspected ovarian cancer. The beauty of the RMI is its simplicity, which has helped it to stand the test of time. Accurate ultrasound assessment of an adnexal mass is fundamental to the success of the RMI. Substituting the subjective assessment by a skilled sonographer35-37 with a score defined by the presence or absence of certain key features facilitates reproducibility. However, the diagnostic test accuracy of the RMI appears lower in external validation studies than the original publications.38, 39 The International Ovarian Tumour Analysis (IOTA) collaboration was formed to encourage the development of better indices by standardising ultrasound reporting across research groups and prospectively collecting a database of features observed in cancerous and benign cysts from which new models could be developed and validated. Initial reports suggest that IOTA models (including simple rules and LR2) demonstrate better sensitivity for the detection of ovarian cancer with a lower false-positive rate than the RMI.38, 40 The challenge is to demonstrate the translation of highly sophisticated ultrasound protocols into ‘real-life’ clinical practice in different centres around the world.41 Women's symptoms have also gained attention as the search for strategies to reduce mortality from ovarian cancer through early detection has gathered pace. To this end, Goff et al.42 developed a symptom index using questionnaire data from women with and without ovarian cancer who were participating in a screening study. Over two-thirds of women had symptoms between 3 and 36 months before diagnosis. Using the Goff index the sensitivity for ovarian disease is 66.7% for a specificity of 90%, which has been validated in a number of external studies.43-45 This has formed the basis of the recommendation in the UK by NICE that symptomatic women are identified as at risk of ovarian cancer and offered further assessment. Much of the criticism surrounding general population screening for ovarian cancer is that low PPV would lead to the overtreatment of large numbers of women. A recent study has demonstrated that the prevalence of ovarian cancer among symptomatic women is ten times higher than that seen in asymptomatic women.46 Furthermore, when offered assessment with CA125 and transvaginal ultrasound at presentation, symptomatic women had lower tumour burdens, which were then more amenable to complete surgical resection, though unfortunately not earlier stage disease than those presenting to gynaecological oncologists by traditional routes. It is suggested that detecting women with lower tumour burdens may account for the increased survival of women diagnosed with stage 3 disease in the first randomised control trials of ovarian cancer screening.47 Data from UKCTOCS showed that the risk of a woman with an adnexal mass developing ovarian cancer within 3 years is 1 in 22.48 Therefore screening asymptomatic women for ovarian cancer will no doubt increase the numbers of women with incidental findings. On the one hand this has offered an unprecedented opportunity to conduct a number of nested cohort studies to assess the natural history of the conditions readily detectable by ultrasonography. But on the other hand, incidental findings cause and will resources in terms of and/or surgery. The of cancers which would not have clinically is a in The incidental finding of cancer is Although has been widely as a risk for cancer in women with postmenopausal the significance of a in the asymptomatic and so its potential use as a screening tool for cancer, is from UKCTOCS suggest that the cut-off for the detection of women with asymptomatic cancer or needs to be in the of as to the used cut-off of in symptomatic But even with an of the risk of cancer in an asymptomatic woman remains low at just Further studies will be needed to the value of early detection of cancer, the acceptability of screening and In the of the roll-out of an ovarian cancer screening that pelvic the need for such studies will more saw a to the cause of ovarian cancer screening. The and Ovarian Cancer Screening which randomised women to ovarian cancer screening, that there was no overall mortality benefit for annual with CA125 and transvaginal ultrasound rate confidence interval Furthermore there was no difference in survival or stage at between the screened and unscreened Additionally, this intervention resulted in more women to from to potential ovarian cancer. in study design between and UKCTOCS provide the that screening may offer for women with ovarian cancer. of the protocol that the study used a single cut-off CA125 level rather than the ROCA to identify women at had clinical in to with positive and follow up for many years after the screening were which time they to Initial data from UKCTOCS have been with of the ovarian cancers identified being as providing evidence for a in screened have suggested that these strategies identify and I cancers which may have clinically apparent within a II cancers representing up to of ovarian cancers diagnosed in symptomatic women, over two-thirds of the ovarian cancers identified by ultrasonography in UKCTOCS were or I screening appears to be more specific for II cancers but screening have had a number of interval cancers all of which were II cancers. These findings are in with the current that ovarian cancers may in the rather than from ovarian the women diagnosed with II cancers, had no adnexal pathology and were diagnosed at an It has been suggested that differences in the that CA125 was used to determine screen in the two trials for the of sensitivity of the trial to detect early Although of the algorithm to the data to any data from the Risk Ovarian Screening Study where women from the general population were using the ROCA followed by transvaginal ultrasound scan if they as high showed specificity and high PPV for the detection of ovarian It is that the UKCTOCS trial will answer whether CA125 as a test will be to translate to in ovarian cancer mortality. If UKCTOCS does not demonstrate survival or mortality benefit from general population ovarian cancer screening, it is to that any randomised trials will be to this given the to such a study The of any benefit from ovarian cancer screening in may be by poor sensitivity of screening to detect true early disease. disease suggests that cancers 90% of the of opportunity where intervention may prognosis at and may have been present for between 8 and years before detectable by current CA125 current screening strategies may detect disease at a that is late to disease The prospective of serum in asymptomatic women may result in the of more relevant of early disease than have been suggested by the comparison of from women with clinically detectable cancers with studies aimed at identifying an from the serum of women who subsequently developed ovarian cancer in UKCTOCS are The years have seen a in the of ovarian cancer, its natural history and the of mortality. Despite in and there has been in the prognosis for women with disease. with the ability of ovarian cancer screening strategies to the of disease to earlier stages, which may translate to better clinical and reduced mortality. screening and ultrasound-based have been shown to be and specific for detecting ovarian cancer but whether these in will provide lead time to the of the disease remains to be success will on the of the multiple and to ensure that research will for better that can be translated into clinical It is that and to the and on the by the high compliance rates seen in the screening studies to work mortality from ovarian cancer. have no to and this was to this is a and is an at the Institute of Cancer University of

The aim of this study was to determine the poly (ADP-ribose) polymerase inhibitors (PARPis) response and outcome of ovarian cancer (OC) patients with BRCA1/2 germline mutation and a history of breast cancer (BC). Thirty-nine OC patients with BRCA1/2 germline mutation and a history of BC were included. The clinicopathological characteristics, PARPis response and prognosis were analyzed. The median interval from BC to OC diagnosis was 115.3 months (range=6.4-310.1). A total of 38 patients (38/39, 97.4%) received platinum-based chemotherapy after surgical removal. The majority of these patients were reported to be platinum sensitive (92.1%, 35/38). 21 patients (53.8%) received PARPis treatment with 16 patients (76.2%) for maintenance treatment and 5 patients (5/21, 23.8%) for salvage treatment. The median duration for PARPis maintenance and salvage treatment was 14.9 months (range=2.0-56.9) and 8.2 months (range=5.2-20.7), respectively. In the entire cohort, 5-year progression-free survival (PFS) and overall survival (OS) rate was 33.1% and 78.9%, respectively. Patients with BRCA1 mutation had a non-significantly worse 5-year PFS (28.6% vs. 45.8%, p=0.346) and 5-year OS (76.9% vs. 83.3%, p=0.426) than those with BRCA2 mutation. In patients with stage III-IV (n=31), first line PARPis maintenance treatment associated with a non-significantly better PFS (median PFS: NR vs. 22.4 months; 5-year PFS: 64.3% vs. 21.9%, p=0.096). The current study shows that these patients may have a good response to platinum-based chemotherapy and a favorable survival. And these patients can benefit from PARPis treatment and will likely be suitable candidates for PARPis.

The role of ultrasound evaluation in the detection of early-stage epithelial ovarian cancer

The United Kingdom Collaborative Trial of Ovarian Cancer Screening (UKCTOCS) recently reported a reduction in the average overall mortality among ovarian cancer patients screened with an annual sequential, multimodal strategy that tracked biomarker CA125 over time, where increasing serum CA125 levels prompted ultrasound. However, multiple cases were documented wherein serum CA125 levels were rising, but ultrasound screens were normal, thus delaying surgical intervention. A significant factor which could contribute to false negatives is that many aggressive ovarian cancers are believed to arise from epithelial cells on the fimbriae of the fallopian tubes, which are not readily imaged. Moreover, because only a fraction of metastatic tumors may reach a sonographically-detectable size before they metastasize, annual screening with ultrasound may fail to detect a large fraction of early-stage ovarian cancers. The ability to detect ovarian carcinomas before they metastasize is critical and future efforts towards improving screening should focus on identifying unique features specific to aggressive, early-stage tumors, as well as improving imaging sensitivity to allow for detection of tubal lesions. Implementation of a three-stage multimodal screening strategy in which a third modality is employed in cases where the first-line blood-based assay is positive and the second-line ultrasound exam is negative may also prove fruitful in detecting early-stage cases missed by ultrasound.

Ovarian cancer is a highly malignant gynecological cancer influenced by the immune microenvironment, metabolic reprogramming, and cellular senescence. This review provides a comprehensive overview of these characteristics. Metabolic reprogramming affects immune cell function and tumor growth signals. Cellular senescence in immune and tumor cells impacts anti-tumor responses and therapy resistance. Targeting immune cell metabolism and inducing tumor cell senescence offer potential therapeutic strategies. However, challenges remain in identifying specific targets and biomarkers. Understanding the interplay of these characteristics can lead to innovative therapeutic approaches. Further research is needed to elucidate mechanisms, validate strategies, and improve patient outcomes in ovarian cancer.

The classification of p53 immunohistochemical staining results and patient outcome in ovarian cancer

Reply: The classification of p53 immunohistochemical staining results and patient outcome in ovarian cancer

Variations in institutional infrastructure, physician specialization and experience, and outcome in ovarian cancer: A systematic review

Biomarkers that predict disease progression might assist the development of better therapeutic strategies for aggressive cancers, such as ovarian cancer. Here, we investigated the role of collagen type XI alpha 1 (COL11A1) in cell invasiveness and tumor formation and the prognostic impact of COL11A1 expression in ovarian cancer. Microarray analysis suggested that COL11A1 is a disease progression-associated gene that is linked to ovarian cancer recurrence and poor survival. Small interference RNA-mediated specific reduction in COL11A1 protein levels suppressed the invasive ability and oncogenic potential of ovarian cancer cells and decreased tumor formation and lung colonization in mouse xenografts. A combination of experimental approaches, including real-time RT-PCR, casein zymography and chromatin immunoprecipitation (ChIP) assays, showed that COL11A1 knockdown attenuated MMP3 expression and suppressed binding of Ets-1 to its putative MMP3 promoter-binding site, suggesting that the Ets-1-MMP3 axis is upregulated by COL11A1. Transforming growth factor (TGF)-beta (TGF-β1) treatment triggers the activation of smad2 signaling cascades, leading to activation of COL11A1 and MMP3. Pharmacological inhibition of MMP3 abrogated the TGF-β1-triggered, COL11A1-dependent cell invasiveness. Furthermore, the NF-YA-binding site on the COL11A1 promoter was identified as the major determinant of TGF-β1-dependent COL11A1 activation. Analysis of 88 ovarian cancer patients indicated that high COL11A1 mRNA levels are associated with advanced disease stage. The 5-year recurrence-free and overall survival rates were significantly lower (P=0.006 and P=0.018, respectively) among patients with high expression levels of tissue COL11A1 mRNA compared with those with low expression. We conclude that COL11A1 may promote tumor aggressiveness via the TGF-β1-MMP3 axis and that COL11A1 expression can predict clinical outcome in ovarian cancer patients.

This study was designed to evaluate the associations between angiogenesis gene polymorphisms and clinical outcome in ovarian cancer patients treated with low-dose cyclophosphamide and bevacizumab. Seventy recurrent/metastatic epithelial ovarian cancer patients were enrolled in a phase II clinical trial. Genomic DNA was available from 53 blood samples. Polymorphisms were analyzed using the PCR-RFLP protocol. A 5' end 33P gammaATP-labeled PCR protocol was used to analyze dinucleotide repeats. Patients genotyped A/A or A/T for the IL-8 T-251A gene polymorphism had a statistically significant lower response rate (19%; 0%) than those homozygous T/T (50%; P = 0.006, Fisher's exact test). Patients carrying a minimum one C allele (C/C; C/T) of the CXCR2 C+785T polymorphism showed a median progression-free survival (PFS) of 7.4 months compared with the PFS of 3.7 months for those homozygous T/T (P = 0.026, log-rank test). Patients with the VEGF C+936T polymorphism C/T genotype had a longer median PFS of 11.8 months, compared with those with the C/C and T/T genotype, which had median PFS of 5.5 months and 3.2 months, respectively (P = 0.061, log-rank test). Patients carrying both AM 3'end alleles < 14 CA repeats had the shortest median PFS of 3.4 months; patients with at least one allele > 14 repeats or both alleles > 14 repeats showed a median PFS of 6.4 months and 7.2 months, respectively (P = 0.008, log-rank test). Our data suggest that the IL-8 A-251T polymorphism may be a molecular predictor of response to bevacizumab-based chemotherapy. The CXCR2 C+785T, VEGF C+936T single nucleotide polymorphisms and the AM 3' dinucleotide repeat polymorphisms may be molecular markers for PFS in ovarian cancer patients.

&lt;div&gt;Abstract&lt;p&gt;&lt;b&gt;Purpose:&lt;/b&gt; This study was designed to evaluate the associations between angiogenesis gene polymorphisms and clinical outcome in ovarian cancer patients treated with low-dose cyclophosphamide and bevacizumab.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Experimental Design:&lt;/b&gt; Seventy recurrent/metastatic epithelial ovarian cancer patients were enrolled in a phase II clinical trial. Genomic DNA was available from 53 blood samples. Polymorphisms were analyzed using the PCR-RFLP protocol. A 5′ end &lt;sup&gt;33&lt;/sup&gt;P γATP-labeled PCR protocol was used to analyze dinucleotide repeats.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Results:&lt;/b&gt; Patients genotyped A/A or A/T for the IL-8 T-251A gene polymorphism had a statistically significant lower response rate (19%; 0%) than those homozygous T/T (50%; &lt;i&gt;P&lt;/i&gt; = 0.006, Fisher's exact test). Patients carrying a minimum one C allele (C/C; C/T) of the CXCR2 C+785T polymorphism showed a median progression-free survival (PFS) of 7.4 months compared with the PFS of 3.7 months for those homozygous T/T (&lt;i&gt;P&lt;/i&gt; = 0.026, log-rank test). Patients with the VEGF C+936T polymorphism C/T genotype had a longer median PFS of 11.8 months, compared with those with the C/C and T/T genotype, which had median PFS of 5.5 months and 3.2 months, respectively (&lt;i&gt;P&lt;/i&gt; = 0.061, log-rank test). Patients carrying both AM 3′end alleles &lt;14 CA repeats had the shortest median PFS of 3.4 months; patients with at least one allele &gt;14 repeats or both alleles &gt;14 repeats showed a median PFS of 6.4 months and 7.2 months, respectively (&lt;i&gt;P&lt;/i&gt; = 0.008, log-rank test).&lt;/p&gt;&lt;p&gt;&lt;b&gt;Conclusion:&lt;/b&gt; Our data suggest that the IL-8 A-251T polymorphism may be a molecular predictor of response to bevacizumab-based chemotherapy. The CXCR2 C+785T, VEGF C+936T single nucleotide polymorphisms and the AM 3′ dinucleotide repeat polymorphisms may be molecular markers for PFS in ovarian cancer patients.&lt;/p&gt;&lt;/div&gt;

5531 Background: Few major studies have examined and compared the management and outcomes of patients from diagnosis to death between countries. We have established an international collaboration across Europe and South Korea to compare treatment and outcomes in Epithelial Ovarian Cancer (EOC). Methods: Patients diagnosed with EOC between January 2012 and December 2018 (age ≥18), were included for analysis. Data from medical records from five European and a single South Korean treatment centre were extracted, standardised to a common data model and analysed at each centre using a common script developed in R. Time to each progression/recurrence event (defined as time to next treatment) and overall survival have been estimated using Kaplan Meier methodology and outcomes stratified by categories of interest. Changes in the use of anti-cancer therapies over time and the incidence of BRCA mutations and incidence/timing of second breast cancers have also been examined. Results: A total cohort of 2925 patients was identified with a median age at each centre of 53 to 67 years. Advanced disease (FIGO stage III - IV) (range 57% to 84%) and high-grade serous morphology (38% to 70%) were most common at each centre. The timing of surgery (primary, interval debulking or delayed) and the proportion of patients undergoing surgery varied with stage. Patients with stage I disease where most likely to undergo surgery (range 73% to 100%) and stage IV the least (range 39% to 84%). Median overall survival for high grade serous cancers ranged from 1.9 to 4.9 years, and for the whole cohort from 2.1 to 5.5 years. Median time to next treatment at first relapse for the whole cohort ranged from 14 to 22 months. Second breast cancers were noted in 6 to 17% of patients and the majority of these occurred before the diagnosis of EOC at a median time of 96 to 118 months prior to diagnosis of EOC. Additional data on treatment pathways, BRCA status and outcomes by line of therapy for each centre will be presented. Conclusions: Preliminary analysis of results across this network suggests a variation in patient populations between sites and substantial differences in both treatments and outcomes. The establishment of a common data model and the use of a common analytic script between sites across six different countries allows for detailed exploration of the factors influencing differences in patient management and treatment outcomes in ovarian cancer patients.

&lt;div&gt;Abstract&lt;p&gt;&lt;b&gt;Purpose:&lt;/b&gt; This study was designed to evaluate the associations between angiogenesis gene polymorphisms and clinical outcome in ovarian cancer patients treated with low-dose cyclophosphamide and bevacizumab.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Experimental Design:&lt;/b&gt; Seventy recurrent/metastatic epithelial ovarian cancer patients were enrolled in a phase II clinical trial. Genomic DNA was available from 53 blood samples. Polymorphisms were analyzed using the PCR-RFLP protocol. A 5′ end &lt;sup&gt;33&lt;/sup&gt;P γATP-labeled PCR protocol was used to analyze dinucleotide repeats.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Results:&lt;/b&gt; Patients genotyped A/A or A/T for the IL-8 T-251A gene polymorphism had a statistically significant lower response rate (19%; 0%) than those homozygous T/T (50%; &lt;i&gt;P&lt;/i&gt; = 0.006, Fisher's exact test). Patients carrying a minimum one C allele (C/C; C/T) of the CXCR2 C+785T polymorphism showed a median progression-free survival (PFS) of 7.4 months compared with the PFS of 3.7 months for those homozygous T/T (&lt;i&gt;P&lt;/i&gt; = 0.026, log-rank test). Patients with the VEGF C+936T polymorphism C/T genotype had a longer median PFS of 11.8 months, compared with those with the C/C and T/T genotype, which had median PFS of 5.5 months and 3.2 months, respectively (&lt;i&gt;P&lt;/i&gt; = 0.061, log-rank test). Patients carrying both AM 3′end alleles &lt;14 CA repeats had the shortest median PFS of 3.4 months; patients with at least one allele &gt;14 repeats or both alleles &gt;14 repeats showed a median PFS of 6.4 months and 7.2 months, respectively (&lt;i&gt;P&lt;/i&gt; = 0.008, log-rank test).&lt;/p&gt;&lt;p&gt;&lt;b&gt;Conclusion:&lt;/b&gt; Our data suggest that the IL-8 A-251T polymorphism may be a molecular predictor of response to bevacizumab-based chemotherapy. The CXCR2 C+785T, VEGF C+936T single nucleotide polymorphisms and the AM 3′ dinucleotide repeat polymorphisms may be molecular markers for PFS in ovarian cancer patients.&lt;/p&gt;&lt;/div&gt;

Introduction: The Hippo signaling pathway controls organ size and inhibits tumorigenesis through a kinase cascade that leads to the phosphorylation of the transcriptional co-activator Yes-associated protein 1 (YAP1, more commonly referred to as YAP). Recent studies have shown that YAP may play a role in the progression of ovarian cancer. However, current reports considering the relationships between YAP expression and clinicopathological outcomes in ovarian cancer are inconsistent. Additionally, the role and functional mechanism of YAP on ovarian tumorigenesis and ovarian cancer progression are not fully understood. Aims: The aims of the present study are to examine the role of YAP in the tumorigenesis of ovarian surface epithelial cells, and to uncover the molecular mechanisms underlying YAP regulation of OSE malignant transformation. Methods: We used a large cohort of ovarian cancer patient samples, an ovarian tissue-specific YAP-overexpression mouse model, and a series of unique cellular models to address above questions. Results: Immunohistochemical analysis using a large cohort of patient samples indicated that YAP expression is associated with poor clinical outcomes in ovarian cancer patients. Overexpression of wild-type YAP or constitutively active YAP in immortalized human ovarian surface epithelial cells (HOSE) induced anchorage-independent growth of these cells in a soft agar assay and tumorigenesis in a xenograft mouse model, suggesting that YAP is sufficient to induce transformation of immortalized human OSEs. To confirm the pathological relevance of our findings, we created an ovarian tissue-specific YAP overexpression mouse model by breeding AMHR2-cre mice to ROSA26-rtTA mice and tetO-YAPS127A mice. The created AMHR2-Cre-rtTA-tetO-YAPS127A mice specifically express YAPS127A (constitutively active YAP) in ovarian tissues once these mice are treated with doxycycline (Dox). We found that AMHR2-induced ovarian tissue specific expression of YAPS127A, which led to drastic neoplasia of ovarian surface epithelial (OSE) cells in AMHR2-Cre-rtTA-tetO-YAPS127A mice after induction with Dox for 3 weeks. In addition, our mechanistic studies indicated that activation of YAP in human OSE cells induced expression of EGFR, ERBB3, HBEGF, NRG1 and NRG2 both in vitro and in vivo. Intriguingly, the activated ERBB signaling in turn suppressed the Hippo pathway and activated YAP protein. Conclusion: Our results demonstrate the existence of a NRGs &amp; HBEGF/ERBBs/YAP/NRGs&amp;HBEGF autocrine loop that may play a critical role in regulating the development and progression of ovarian cancer. Our study clearly indicates that combined targeting of ERBB and the Hippo/YAP pathways represents a novel strategy for treatment of ovarian cancer. Citation Format: Chunbo He, Xiangmin LV, John S. Davis, Wang Cheng. YAP interacts with the ERBB signaling pathway to control ovarian surface epithelial cell tumorigenesis. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: Exploiting Vulnerabilities; Oct 17-20, 2015; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(2 Suppl):Abstract nr A29.