SIVA-1 interaction with PCBP1 serves as a predictive biomarker for cisplatin sensitivity in gastric cancer and its inhibitory effect on tumor growth in vivo.

Background: SIVA-1 has been reported to play a key role in cell apoptosis and gastric cancer (GC) chemoresistance in vitro. Nevertheless, the clinical significance of SIVA-1 in GC chemotherapy remains unclear. Methods and results: Immunohistochemistry and histoculture drug response assays were used to determine SIVA-1 expression and the inhibition rate (IR) of agents to GC and to further analyze the relationship between these two phenomena. Additionally, cisplatin (DDP)-resistant GC cells were used to elucidate the role and mechanism of SIVA-1 in vivo. The results demonstrated that SIVA-1 expression was positively correlated with the IR of DDP to GC but not with those of 5-fluorouracil (5-FU) or adriamycin (ADM). Furthermore, SIVA-1 overexpression with DDP treatment synergistically inhibited tumor growth in vivo by increasing PCBP1 and decreasing Bcl-2 and Bcl-xL expression. Conclusions: Our study demonstrated that SIVA-1 may serve as an indicator of the GC sensitivity to DDP, and the mechanism of SIVA-1 in GC resistance to DDP was preliminarily revealed.

Efficacy of Recombinant Methioninase on Late-stage Patient Cancer in the Histoculture Drug Response Assay (HDRA) as a Potential Functional Biomarker of Sensitivity to Methionine-restriction Therapy in the Clinic.

Background/Aim: The present study utilized the three-dimensional histoculture drug response assay (HDRA) to determine the efficacy of recombinant methioninase (rMETase) on tumor tissue resected from patients with late-stage cancer, as a functional biomarker of sensitivity to methionine restriction therapy. Patients and Methods: Resected peritoneal-metastatic cancer, including colorectal cancer, pancreatic cancer, ovarian cancer, and pseudomyxoma were placed on Gelform in RPMI 1640 medium for seven days and treated with rMETase from 2.5 U/ml to 20 U/ml. Cell viability was determined using the MTT assay. A total of 48 patients with late-stage cancer underwent testing for rMETase responsiveness using the HDRA. Results: Colorectal cancer and pseudomyxoma had the highest sensitivity to rMETase. Pancreatic and ovarian cancer also responded to rMETase, but to a lesser degree. Conclusion: Patients with tumors with at least 40% sensitivity to rMETase in the HDRA are being considered as candidates for methionine restriction therapy, which includes the use of rMETase in combination with a low-methionine diet.

CHFR-Promoter-Methylation Status Is Predictive of Response to Irinotecan-based Systemic Chemotherapy in Advanced Colorectal Cancer.

BACKGROUND/AIM: We investigated whether promoter methylation of the checkpoint-with-forkhead-and-ring-finger-domains (CHFR) gene is a predictor of the efficacy of irinotecan-based systemic chemotherapy for advanced colorectal cancer (CRC) patients. MATERIALS AND METHODS: CHFR-promoter methylation was measured by quantitative methylation-specific PCR (qMSP). The histoculture drug response assay (HDRA) was used in vitro to analyze the correlation between CHFR-promoter methylation and the efficacy of the irinotecan-active-metabolite SN38 in colorectal-cancer tissues from 44 CRC patients. CHFR promoter-methylation was also analyzed for its correlation with clinical response to irinotecan-based systemic chemotherapy of 49 CRC patients. RESULTS: CHFR-promoter methylation significantly-positively correlated with inhibition of colon cancer by SN38 in the HDRA (p=0.002). CHFR-promoter methylation also significantly-positively correlated with clinical response to irinotecan-based systemic chemotherapy (p=0.04 for disease control). CHFR-promoter methylation also significantly-positively correlated (p=0.01) with increased progression-free survival for patients treated with irinotecan-containing FLOFIRI in combination with bevacizumab, the most-frequent regimen in the cohort. CONCLUSION: Sensitivity of advanced CRC patients to irinotecan-based systemic chemotherapy can be predicted by the extent of CHFR-promoter methylation.

Clinical Correlation of the Histoculture Drug Response Assay in Gastrointestinal Cancer.

The histoculture drug response assay (HDRA) with tumors histocultured on Gelfoam® was tested for clinical correlation for advanced gastric and colorectal cancer patients. In one study, 29 patients were treated with drugs shown to be ineffective in the HDRA, and all 29 cases showed clinical chemoresistance. In nine patients treated with drugs shown to be effective in the HDRA, six showed clinical chemoresponse and three showed arrest of disease progression. In a study of 32 patients with stage III and IV gastric cancer treated with mitomycin C and 5-fluorouracil (5-FU), the survival rate of 10 patients whose tumors were sensitive to either mitomycin C and/or 5-fluorouracil in the HDRA was significantly better than that of 22 patients whose tumors were insensitive to both drugs in the HDRA. Twenty-nine patients with stage III and IV colorectal cancer without remaining measurable tumor lesions after surgery were treated with fluoropyrimidines adjuvantly. The recurrence-free survival rate of 7 patients whose tumors were sensitive to 5-fluorouracil in the HDRA was significantly better than that of 22 patients whose tumors were insensitive in the HDRA. In a companion study of 128 gastric cancer patients whose tumors were evaluated in the HDRA, the overall and disease-free survival rates of the HDRA-sensitive group were found to be significantly higher than those of the HDRA-resistant group, treated with the same drugs.

Prospective Clinical Correlation of the Histoculture Drug Response Assay for Ovarian Cancer.

The histoculture drug response assay (HDRA) has been correlated clinically to a number of cancer types (please see Chaps. 7 - 11 of the present volume). The present chapter reviews the clinical trials of the HDRA for ovarian cancer. A prospective clinical trial of the HDRA for advanced epithelial ovarian cancer (AEOC) was performed at Asan Medical Center, Seoul, Korea. The clinical trial compared the efficacy of first-line therapy paclitaxel and carboplatinum in the HDRA and the clinical response for the patients whose tumors were tested in the HDRA. A series of patients (104) were treated with adjuvant combination chemotherapy of paclitaxel and carboplatinum after primary cytoreductive surgery. Tumor fragments were cultured on Gelfoam® and tested with paclitaxel and carboplatinum and evaluated with the MTT endpoint. Patients were categorized into two groups as either sensitive to both drugs (SS) or not sensitive to one or both drugs (R) based on HDRA results. The recurrence rate was much lower in the SS group compared to the R group, 29.2% vs. 69.8%, respectively. The SS group had a significantly longer progression-free survival compared to the R group, 34.0 months vs. 16.0 months, respectively. In another clinical trial, the HDRA was performed on 85 cases of ovarian cancer and 97% were evaluable. HDRA results were correlated to clinical response of 15 patients who received cisplatinum-based therapy that included doxorubicin and cyclophosphamide (CAP therapy). The true-positive rate was 88%, the true-negative rate was 86%, the sensitivity was 88%, the specificity was 86%, and the accurate prediction rate was 87% when HDRA results were compared to the response of the treated patients.

Clinical Correlation of the Histoculture Drug Response Assay for Head and Neck Cancer.

Gelfoam® histoculture was utilized to develop the histoculture drug response assay (HDRA) for head and neck cancer. Specimens of head and neck tumors were evaluated for sensitivity to the following drugs: cisplatinum (CDDP), 5-fluorouracil (5-FU), and the combination of CDDP and 5-FU. In the first clinical study at UCSD, 10 of 12 patients with tumors that were drug sensitive in Gelfoam® histoculture had either complete or partial response clinically. Comparisons of HDRA results, obtained with [3H]thymidine incorporation as the endpoint were made with clinical responses, i.e., complete response, partial response, or no response. The overall accuracy of the HDRA was 74% in this correlative clinical trial; the predictive positive value was 83%, the sensitivity was 71%, and the specificity was 78%. Seven of 11 patients with HDRA-resistant tumors demonstrated no response for a predictive negative value of 64%. In a subsequent study at Memorial Sloan Kettering Cancer Center, tumor specimens from 41 to 42 patients undergoing treatment for head and neck cancer were successfully evaluated by the HDRA. The histocultured tumors were treated with 5-FU and/or CDDP and a control group received no drug treatment. After completion of drug treatment, the relative cell survival in the tumors was determined using the MTT endpoint. Sensitivity was defined as a tumor inhibition rate (IR) of greater than 30%. Survival comparisons were performed using the generalized Wilcoxon test for the comparison of Kaplan-Meier survival curves. Resistance to 5-FU was observed in 13 cases (32%), to CDDP in 13 cases (32%), and to both agents in 11 cases (27%). The 2-year cause-specific survival was significantly greater for patients sensitive to 5-FU than patients who were resistant (85% vs. 64%), CDDP (86% vs. 64%), or both agents (85% vs. 63%). These results demonstrate the clinical usefulness of the HDRA for head and neck cancer.

Clinical Usefulness of the Histoculture Drug Response Assay for Breast Cancer.

The histoculture drug response assay (HDRA) was used to compare drug sensitivity of recurrent and primary breast cancer in vitro as well as in the clinic. The HDRA utilizes 3-dimensional culture of human tumors on Gelfoam®. The evaluation rate was 98.8%. The HDRA mean inhibition rate of primary tumors vs. recurrent tumors was, respectively, 57.9% and 38.6% for doxorubicin (DOX); 59.9% and 42.8% for mitomycin C (MMC); 49.0% and 33.4% for 5-fluorouracil (5-FU); and 34.5% and 16.0% for cisplatinum (CDDP). The recurrent cases were pretreated clinically with CAF (cyclophosphamide [CTX], DOX, and 5-FU), CEF (CTX, epirubicin [EPN], and 5-FU), or CMF (CTX, methotrexate [MTX], and 5-FU). 64.7% of the recurrent cases were resistant to all four agents tested compared to 27% of the primary cases. Only 5.9% of the recurrent cases were sensitive to three or more agents as opposed to 18% of the primary cases. The correlation of the HDRA results to clinical outcome in another breast-cancer study was 80.0% with 15 cases evaluated consisting of five true positives, three false positives, seven true negatives, and no false negatives. HDRA was also performed on surgical specimens of primary tumor and axillary lymph node metastasis from each of 30 breast cancer patients. The lymph-node metastases were more resistant than the primary tumor for DOX, 5-FU, and MMC, but not for CDDP. The data suggest that both primary tumor and metastases from individual patients should be tested in the HDRA to enhance clinical efficacy of chemotherapy. There also was a lack of correlation with breast cancer subtype and drug response in the HDRA, further suggesting the importance of individualized treatment for breast cancer patients afforded by the HDRA.

Application of drug sensitivity from three-dimensional tissue in individualized precision medical treatment for tumor / 实用肿瘤学杂志

At present,most chemotherapy schemes for malignant tumors use the standard chemotherapy regimen recommen-ded by the National Comprehensive Cancer Network(NCCN)clinical practice guidelines,because they ignore the inherent heterogenei-ty of the tumor,resulting in low efficiency,high toxic and side effects,and high costs issues. Therefore,the realization of " individual-ized and accurate medical care" for cancer is a general trend. The sensitivity screening of chemotherapy drugs for cancer patients to a-chieve individualized precise drug delivery is one of the main contents of "individualized precision medicine". The three-dimensional histoculture drug response assay( HDRA) is a method for detecting drug sensitivity after in vitro cultivation of active tumor tissue blocks obtained by surgical resection or biopsy. It not only has a short experimental cycle,but it also maintains the tumor tissue struc-ture,heterogeneity and micro-environment,which have high clinical practice consistency,and it is a relatively promising technique for detecting drug susceptibility. Therefore,this article reviews the development history,clinical application and the future development trend of HDRA.

Independence of cytotoxic drug sensitivity profiles and receptor subtype of invasive ductal breast carcinoma demonstrated by the histoculture drug response assay (HDRA).

AIM: Breast cancer can be divided into four subtypes: luminal-A, luminal-B, HER-2 enriched and triple negative breast cancer (TNBC) by the presence or absence of receptors. Each subtype has a typical clinical behavior and prognosis. Many chemotherapeutic agents are used clinically for breast cancer. The histoculture drug response assay (HDRA) is used for selection of effective chemotherapeutic agents for individual patients Materials and Methods: In the present study, the HDRA was used for eleven frequently-used single-agent or combinations on the four subtypes of breast cancer in order to determine the correlation of drug sensitivity profile and breast-cancer subtype. Fifty invasive ductal breast carcinoma patients who underwent cancer surgery and adjuvant chemotherapy between January 2012 and January 2013 had their tumors analyzed in the HDRA. Age, gender, height and weight, tumor-nodes-metastasis (TNM) stage, immunohistochemical (IHC profiles, breast-cancer subtypes and HDRA results were recorded. RESULTS: The inhibition rate (IR) of each agent or combination for each breast-cancer subtype was determined. Drug to drug IRs were statistically distinct in all subtypes (p<0.05) but no correlation between response to chemotherapeutic agents and breast-cancer subtype was found (p=0.851 by two-way ANOVA test). CONCLUSION: The clear difference between average sensitivity of the chemotherapeutic agents tested and lack of correlation with breast-cancer subtype suggest the importance of individualized treatment for breast-cancer patients.

Can the Histoculture Drug Response Assay Predict the Clinical Results of Chemotherapy in Breast Cancer? / 한국유방암학회지

PURPOSE: The behavior of invasive carcinomas in human can be very varied with different individual responses to chemotherapy. Individualization is crucial to the optimization of chemotherapy. Therefore, the prediction of a tumor's sensitivity to anticancer agents has been the subject of intensive investigation. In order to investigate the pathobiology of breast cancer, it is necessary to maintain or recreate the characteristics of the three-dimensional architecture of the tissues in culture. In this study, we have evaluated the relationship between the Histoculture Drug Response Assay (HDRA) assessment and chemotherapy responses in breast cancer patients. METHODS: Tumor specimens from 30 patients with breast cancer were evaluated using the HDRA. Tumor tissues were cultured on gelfoam sponge gel in 24-well plates, followed by treatment with a variety of chemotherapeutic agents. All treatments were conducted in triplicate. The sensitivity of a chemotherapy regimen was defined as a tumor inhibition rate (IR) in excess of 30%. Neoadjuvant or palliative chemotherapy for patients, using anthracycline or taxane, was conducted on the basis of the established protocols. The responses to treatments were compared with the results of the HDRA. RESULTS: The mean IR for the combinations of doxorubicin and docetaxel and for FAC and AC were 48, 45, and 36%, respectively. The above partial rate of response to chemotherapy was 81.1%. The sensitivity and specificity of the HDRA assessment, with a 30% inhibition rate, were 81.5 and 66.7%, respectively. The positive and negative response prediction values were 91.7 and 44.4%, respectively. The responses to treatments and the results of the HDRA assessment were not correlated with the expressions of the hormonal receptor or c-erbB2. CONCLUSION: In cases in which the inhibition rate is in excess of 30%, the HDRA assessment yielded a high positive response prediction value. The sensitivity to chemotherapy, as determined by the HDRA, appears to be a good guide for selection in breast cancer patients. Thus the results presented herein should be integrated into future research on the subject.

Can the Histoculture Drug Response Assay Predict the Clinical Results of Chemotherapy in Breast Cancer? / 한국유방암학회지

PURPOSE: The behavior of invasive carcinomas in human can be very varied with different individual responses to chemotherapy. Individualization is crucial to the optimization of chemotherapy. Therefore, the prediction of a tumor's sensitivity to anticancer agents has been the subject of intensive investigation. In order to investigate the pathobiology of breast cancer, it is necessary to maintain or recreate the characteristics of the three-dimensional architecture of the tissues in culture. In this study, we have evaluated the relationship between the Histoculture Drug Response Assay (HDRA) assessment and chemotherapy responses in breast cancer patients. METHODS: Tumor specimens from 30 patients with breast cancer were evaluated using the HDRA. Tumor tissues were cultured on gelfoam sponge gel in 24-well plates, followed by treatment with a variety of chemotherapeutic agents. All treatments were conducted in triplicate. The sensitivity of a chemotherapy regimen was defined as a tumor inhibition rate (IR) in excess of 30%. Neoadjuvant or palliative chemotherapy for patients, using anthracycline or taxane, was conducted on the basis of the established protocols. The responses to treatments were compared with the results of the HDRA. RESULTS: The mean IR for the combinations of doxorubicin and docetaxel and for FAC and AC were 48, 45, and 36%, respectively. The above partial rate of response to chemotherapy was 81.1%. The sensitivity and specificity of the HDRA assessment, with a 30% inhibition rate, were 81.5 and 66.7%, respectively. The positive and negative response prediction values were 91.7 and 44.4%, respectively. The responses to treatments and the results of the HDRA assessment were not correlated with the expressions of the hormonal receptor or c-erbB2. CONCLUSION: In cases in which the inhibition rate is in excess of 30%, the HDRA assessment yielded a high positive response prediction value. The sensitivity to chemotherapy, as determined by the HDRA, appears to be a good guide for selection in breast cancer patients. Thus the results presented herein should be integrated into future research on the subject.

In Vitro Chemosensitivity Test of Human Soft Tissue Sarcoma Using the HDRA (Histoculture Drug Response Assay) Method / 대한정형외과학회잡지

PURPOSE: To investigate the variation in the chemosensitivity in soft tissue sarcoma (STS), fresh biopsy with sample for culture was tested using the histoculture drug response assay (HDRA)method. MATERIALS AND METHODS: 30 samples of fresh STS were obtained during either biopsy or surgical removal at our hospital between March, 2002 and March, 2004. RESULTS: Drug sensitivity testing by HDRA showed that two drug, Doxorubicin and CDDP, had a significantly higher inhibition rate than BLM, CTX, DTIC, VCR or VP-16 in the thirty STS tested. Doxorubicin showed the highest inhibition rate in the liposarcoma. CDDP shoewd the significant inhibition rate in the synovial sarcoma and malignant fibrous histiocytpma. Depending on the morphological type, round cell sarcoma and pleomorphic sarcoma were more sensitive to Doxorubicin and CDDP than spindle cell sarcoma. In the round cell sarcoma, BLM, CTX, VP-16 and IFS also showed above 30% inhibition rate. CONCLUSION: Drug sensitivity testing in STS should be evaluated with clinical outcome in the future and then HDRA will provide useful information for selection of an anticancer agent for STS because of its ease of evaluation and high predictability.

Histoculture Drug Response Assay in Colorectal Cancer Specimen

PURPOSE: For decades, systemic medical treatment for colorectal cancer has been limited almost entirely to 5- fluorouracil (5-FU). In cases of advanced colorectal cancer, the response rate to standard 5-FU regimen was lower than 20%. Recently, new drugs that have another action mechanism have been introduced-irinotecan, oxaliplatin, raltitrexed, capecitabine, etc. Clinicians have to choose the appropriate drug for advanced cases. Until recently, choice of chemotherapeutic agents was based on the experience of clinicians, or on retrospective or prospective clinical trial reports. In this study, we performed HDRA (histoculture drug response assay) to assess the effectiveness of chemotherapeutic agents in colorectal cancer. METHODS: Tumor specimens of 33 colorectal cancers were collected in 15 ml tubes containing PBS buffer. Tissues were minced using an autoclaved knife and histocultured on collagen sponge gel matrix, followed by treatment with 5-FU, 5-FU & leucovorin, oxaliplatin, oxaliplatin & 5-FU, irinotecan, or irinotecan & 5-FU. After 48 hours, cell viability was assessed by MTT assay. The inhibition rate of each drug was calculated for relative survival. Cases of drug responsibility below 30% were regarded as drug resistant cases. RESULTS: Thirty cases were tested. Three cases had synchronous lesion. Thirty-three tissues were evaluated using HDRA. Seventeen cases (53.3%) were rectal cancer. The initial 6 cases were tested using a single agent the other 27 cases were tested using combined agents. The regimen showing the best responses was oxaliplatin with 5-FU (8/26 cases, 30.8%). Seven cases were regarded as chemoresistant cases because they showed low IR below 30% for all agents. Synchronous lesions showed similar drug responses. CONCLUSION: HDRA is relatively simple and easily applicable to in vitro study to determine the appropriate chemotherapeutic agents. Further study is necessary to assess the effectiveness including tumor recurrence and survival.

Correlation of Immunohistochemical Expression of MDR1, MRP1, Topoisomerase IIalpha with Prognostic Factors and Histoculture Drug Response Assay (HDRA) Result in Breast Carcinoma / 한국유방암학회지

PURPOSE: Drug resistance plays an important role in the failure of chemotherapy in breast cancer. The purpose of the study was to investigate the chemosensitive and chemoresistance indices of breast carcinomas and see if the in vitro chemosensitivity test correlated with the prognostic indices. METHODS: The immunohistochemical expressions of MDR1, MRP1 and topoisomerase IIalpha(topo IIalpha) were studied and then correlated these with the in vitro chemosensitivities using the histoculture drug response assay (HDRA) and clinicopathological factors in 51 breast carcinomas. RESULTS: In the breast carcinomas examined, the immunohistochemical expressions of MDR1, MRP1 and topo II alpha were strongly observed in 26 (51.0%), 16 (32.0%), 15 (31.3%) carcinomas, respectively. The MRP1 was more frequently expressed in poorly differentiated carcinomas (P= 0.006), and those of MDR1 and topo II alpha were more frequently observed in tumor overexpressing cerbB2 (P=0.038, P=0.036). The expression of MDR1 was related to that of topo II alpha (P=0.015). Comparing these markers with the in vitro chemosensitivities to cyclophosphamide, 5-FU, adriamycin, taxol and taxotere, no correlations were found between the expression of MDR1, MRP1, and topo II alpha but from the chemosensitivity using the HDRA, the growth inhibition rate for cyclophosphamide was higher in MRP1 expressing carcinomas (P=0.009). CONCLUSION: MDR1, MRP1 and topo II alpha were all found to be associated with the poor prognostic indices, but assessment of their immunohistochemical expressions did not allow for prediction of the response to chemotherapy by the in vitro chemosensitivity test in breast carcinomas.