Harukaze Yamamoto

64Cu-DOTA-Trastuzumab PET Imaging in Patients with HER2-Positive Breast Cancer

The purpose of this study was to determine the safety, distribution, internal dosimetry, and initial human epidermal growth factor receptor 2 (HER2)–positive tumor images of 64Cu-DOTA-trastuzumab in humans. Methods: PET was performed on 6 patients with primary or metastatic HER2-positive breast cancer at 1, 24, and 48 h after injection of approximately 130 MBq of the probe 64Cu-DOTA-trastuzumab. Radioactivity data were collected from the blood, urine, and normal-tissue samples of these 6 patients, and the multiorgan biodistribution and internal dosimetry of the probe were evaluated. Safety data were collected for all the patients after the administration of 64Cu-DOTA-trastuzumab and during the 1-wk follow-up period. Results: According to our results, the best timing for the assessment of 64Cu-DOTA-trastuzumab uptake by the tumor was 48 h after injection. Radiation exposure during 64Cu-DOTA-trastuzumab PET was equivalent to that during conventional 18F-FDG PET. The radioactivity in the blood was high, but uptake of 64Cu-DOTA-trastuzumab in normal tissues was low. In 2 patients, 64Cu-DOTA-trastuzumab PET showed brain metastases, indicative of blood–brain barrier disruptions. In 3 patients, 64Cu-DOTA-trastuzumab PET imaging also revealed primary breast tumors at the lesion sites initially identified by CT. Conclusion: The findings of this study indicated that 64Cu-DOTA-trastuzumab PET is feasible for the identification of HER2-positive lesions in patients with primary and metastatic breast cancer. The dosimetry and pharmacologic safety results were acceptable at the dose required for adequate PET imaging.

Molecular imaging using PET for breast cancer

Molecular imaging can visualize the biological processes at the molecular and cellular levels in vivo using certain tracers for specific molecular targets. Molecular imaging of breast cancer can be performed with various imaging modalities, however, positron emission tomography (PET) is a sensitive and non-invasive molecular imaging technology and this review will focus on PET molecular imaging of breast cancer, such as FDG-PET, FLT-PET, hormone receptor PET, and anti-HER2 PET.

Phase I and pharmacokinetic study of trastuzumab emtansine in Japanese patients with HER2-positive metastatic breast cancer

OBJECTIVE Trastuzumab emtansine (T-DM1), an antibody-drug conjugate composed of the cytotoxic agent DM1 conjugated to trastuzumab via a stable thioether linker, has shown clinical activity in human epidermal growth factor receptor 2-positive metastatic breast cancer patients. This study evaluated the maximum tolerated dose, toxicity and pharmacokinetics of trastuzumab emtansine in Japanese breast cancer patients. METHODS Inoperable advanced or recurrent human epidermal growth factor receptor 2-positive breast cancer patients were administered trastuzumab emtansine intravenously at a dose of 1.8, 2.4 or 3.6 mg/kg every 3 weeks. The maximum tolerated dose was estimated using the continual reassessment method. RESULTS This study enrolled 10 patients who were administered trastuzumab emtansine for a median of seven cycles. The dose-limiting toxicity was Grade 3 elevation of aspartate aminotransferase/alanine aminotransferase at the 2.4 mg/kg dose level. The maximum tolerated dose was estimated to be 3.6 mg/kg because at the point when dose-limiting toxicity was evaluable in 10 patients, the probability of dose-limiting toxicity estimated using the continual reassessment method was closest to 25% at a dose of 3.6 mg/kg and this was unchanged by the results for patients enrolled after that. The most frequent adverse events were nausea, arthralgia, fever, fatigue and decreased appetite. Adverse events were generally tolerable. The maximum concentration and area under the concentration-time curve increased linearly with the dose. CONCLUSIONS Trastuzumab emtansine up to 3.6 mg/kg was well tolerated by Japanese breast cancer patients. Although thrombocytopenia and hepatotoxicity tended to be more severe than was seen in Western patients in previous trastuzumab emtansine trials, those adverse events recovered without special supportive treatment.

Use of squamous cell carcinoma antigen as a biomarker of chemotherapy response in patients with metastatic cervical carcinoma

OBJECTIVE To examine the use of squamous cell carcinoma antigen (SCCA) as a biomarker of chemotherapy response in patients who underwent chemotherapy for metastatic cervical carcinoma. STUDY DESIGN The study population consisted of patients who underwent first-line chemotherapy for metastatic cervical carcinoma between 1999 and 2009. SCCA levels were serially measured before, during and after chemotherapy. Radiographic responses were evaluated according to the criteria of the World Health Organization. A logistic model was used to determine the best prediction model, and internal and external validation of the prediction model were performed to compare the areas under the receiver operating characteristic curves (AUCs). RESULTS In total, 55 patients were included in the analysis. Data for 32 patients enrolled in various clinical trials were used to develop the prediction model. Patients who achieved a radiographic response showed a significant decline in SCCA levels between the second and third cycles of chemotherapy, whereas patients who did not achieve a radiographic response showed constant SCCA levels over the same period. The prediction model was developed on the basis of changes in the SCCA level between the second and third cycles of chemotherapy (AUC=0.832) and the baseline SCCA level. The AUC after external validation, calculated using the data of the clinical practice population (n=22), was 0.871. CONCLUSIONS A response to chemotherapy was possible for patients in whom SCCA levels declined between the second and third cycles of chemotherapy.

Fc-Gamma receptor polymorphism and gene expression of peripheral blood mononuclear cells in patients with HER2-positive metastatic breast cancer receiving single-agent trastuzumab

AimThe aim of this study was to investigate gene expression in the peripheral blood mononuclear cells (PBMCs) of patients with HER2-positive breast cancer receiving trastuzumab. We also evaluated the effect of Fc-gamma receptor genotype on trastuzumab-driven gene expression.Materials and methodsGene expression was assessed by microarray analyses before and after administration of single-agent trastuzumab in 34 patients with metastatic HER2-positive breast cancer who were genotyped for Fc-gamma receptor (FcGR) IIA H131R and FcGRIIIA V158F. Gene set enrichment analysis (GSEA) was used to identify the gene sets that were significantly enriched after administration of trastuzumab in patient cohorts categorized by FcGR variant.ResultsAt baseline three non-immune-related gene sets were identified only in patient cohort of FcGRIIA non-H/H variant. Thirty gene sets were identified in the cohort of FcGRIIIA V/V variants, while no gene set was identified in FcGRIIIA non-V/V variants one week after starting trastuzumab. Eleven gene sets were identified in FcGRIIA H/H variants 8 week after starting trastuzumab, but none in non-H/H variants. Immune-related gene sets were significantly down-regulated after administration of trastuzumab.ConclusionThe response of PBMCs to trastuzumab markedly varied with polymorphisms in FcGRIIA and FcGRIIIA. These results indicate that FcGR polymorphisms contribute to the systemic immune reaction triggered by trastuzumab. Further investigations are needed to clarify the biological effects of FcGR variation on the mechanism of trastuzumab activity.

A descriptive analysis of post-chemotherapy development of interstitial lung disease using spontaneous reporting data in Japan.

This descriptive study used the Japanese spontaneous reporting data to investigate the time taken (TTILD) to development of interstitial lung disease (ILD) after initiation of chemotherapy and the death rates attributed in part to post-chemotherapy ILD (i.e., DR) for anticancer drugs. We evaluated TTILD and DR endpoints for 36 anticancer drugs, which are widely used for treating 11 solid and 3 hematological cancers, and are suspected of causing ILD, by using 8- year spontaneous reporting data recording for 2,553 patients in the reporting system of the relevant Japanese regulatory agency. The median TTILD and overall DR attributable to post-chemotherapy ILD for the drugs were 1.8 months and 29%, respectively. For most drugs, the median TTILDs were between 1 to 4 months, and the DRs attributable to postchemotherapy ILD were <40%; however, TTILDs were as long as 4 to 6 months and DRs attributable to postchemotherapy ILD were ≥40% for several other drugs. Of the 36 drugs, we identified those that may trigger postchemotherapy late-onset ILDs or result in high DRs. The anticancer drugs that may have triggered late-onset ILDs were defined as those that caused ILD development after approximately 4 months from the initial drug administration.

441OIMAGING MASS SPECTROMETRY OF NOVEL DRUG IN HUMAN TUMOR SPECIMENS: DISTRIBUTION OF UNLABELED DRUGS TO SUPPORT EARLY PHASE CLINICAL TRIAL

ABSTRACT Aim: Assessment of drug pharmacokinetics is an important component of early phase drug development. Imaging Mass Spectrometry (IMS) is an innovative technique in the preclinical study that allows for analysis of the distribution of target molecules in tumor tissues. The advantage of imaging technology is the detection of the molecule of interest in tissues without labeling. We performed tumor biopsies in patients with solid tumors who were participating in a phase I trial (NCT01813474) of olaparib. The aim of this study was to examine drug distribution in the tumor biopsy specimens by IMS, which provides a sensitive and label-free approach to imaging drugs. Methods: Patients with solid tumors received the tablet formulation of olaparib in dose escalation (200mg BID; 300mg BID) and expansion (300mg BID) cohorts. The timing of biopsies in consenting patients was during cycle 2 and/or at the time of progression. IMS was performed using an Imaging Mass Microscope (Shimadzu, Japan). The concentrations of olaparib in tissues were validated by using LC-MS/MS. Results: In total, seven tumor biopsies were performed in six patients with solid tumors; three breast cancer including one BRCA1 mutation positive, one ovarian cancer, one peritoneal cancer, one cervical cancer. One patient had biopsies performed at the time of drug administration and progression. One patient received olaparib 200mg BID; the remaining patients received olaparib 300mg BID. IMS signal levels of olaparib correlated well with the concentration of drug in tumor tissues derived from patients using LC-MS/MS, a conventional method used in pharmacokinetic studies. The distribution of Olaparib was in the tumor region and the signal level in areas of necrosis was higher than that observed in living cell areas. Conclusions: The use of IMS has allowed to follow the distribution of an unlabeled olaparib in target tissues. In addition, this technique can also allow further understanding of PK/PD relationships of olaparib in combination with other compound at clinical trial. Disclosure: All authors have declared no conflicts of interest.

1575PINTEGRATIVE ANALYSIS OF TWO PROSPECTIVE NEOADJUVANT STUDIES WITH BREAST CANCER PATIENTS AND MICROARRAY ANALYSIS

ABSTRACT Aim: Two prospective studies were designed to examine the feasibility of gene expression profiling to predict pathological response in early breast cancers. We present long-term outcome data as well as association of gene expression with clinical data. Methods: Clinical characteristics of two consecutive prospective phase II neoadjuvant studies were examined. Eligible criteria included stage IIA-IIIC, chemotherapy-naive, measurable disease, age >20, PS 0/1, adequate organ function. Neoadjuvant treatment included anthracyclin-based regimens plus taxane with or without trastuzumab. cDNA microarray by Affimetrix Gene Chip U133 plus 2.0 arrays with 54613 genetic variables from pretreatment fine-needle biopsy specimens was performed. Results: Between December 2005 and December 2013, 107 patients were enrolled and followed up. Median follow up time was 50.8 months (13.2-92.7). Median age 51 (23-76), PS 0:1=100:7, stages IIA:IIB:IIIA:IIIB:IIIC=23:50:19:14:1, HER2-/ER+:HER2+/ER+:HER2+/ER-:HER2-/ER-=48:13:23:23. Pathological complete response (pCR) rate was 28%. pCR rate was significantly higher in HER2+ plus ER+, HER2+ plus ER-, and HER2- plus ER- breast cancers compared to HER2- plus ER+ cancers (p=0.0006, p=0.013, p=0.0218 respectively), and lymph node (LN) negative cancers compared to LN 1-3 positive cancers (p=0.0098). Five year disease free survival (DFS) was 77.7% (HER2-/ER+:HER2+/ER+:HER2+/ER-:HER2-/ER-= 75.8%:92.3%:86.7%:60.9%). Significant prolonged DFS was seen in HER2-/ER+ cancers compared to HER2-/ER- (p=0.0008), and LN negative cancers compared to LN>=4 (p=0.0025). No significance was seen between DFS and pCR. Total cDNA microarray data of 78 of 107 patients was available. For DFS, 17 most-associated genes were identified. The associated genes were evaluated using Cox regression, and low- risk classification identified patients (n=46) showed improved DFS than the high-risk group (n=33) (HR 70.0, p=0.004). For pCR, 8 specific genes were identified as having a relation. Conclusions: Clinical characteristics from long term follow up of two prospective studies were shown. Microarray analysis enabled us to identify preliminary data of specific genes associated with breast cancer recurrence and pCR respectively. Further clinical follow-up and validation tests are to follow. Disclosure: All authors have declared no conflicts of interest.

379PRISK FACTORS FOR DEVELOPING SKELETAL-RELATED EVENTS ASSOCIATED WITH METASTATIC BREAST CANCER PATIENTS RECEIVING BONE-MODIFYING AGENTS

ABSTRACT Aim: Bone-modifying agents (BMAs) reduce the incidence of skeletal-related events (SREs) and are recommended for breast cancer patients with bone metastasis. However, the risk factors for developing SREs, during BMA treatment, are not adequately studied. This study evaluated the time to first SRE and the time between first and multple SREs in order to identify the factors predicting SREs. Methods: The medical records of 534 women with breast cancer who developed bone metastasis, between 1999 and 2011, were reviewed. SREs were defined as a pathologic fracture, spinal cord compression, or the necessity of bone irradiation or surgery. The factors investigated included the type of primary cancer, performance status, menopause status, presence of extraskeletal metastasis, prior history of bone fractures and palliative radiation therapy, serum levels of lactate dehydrogenase and calcium corrected for albumin level at the time of the initial BMA dose, BMA type, and time between the first diagnosis of bone metastasis and the initiation of BMAs (0-6 months vs. above 6 months). These factors were analyzed using Cox proportional hazard analyses and Andersen-Gill approach. Results: Multivariate analyses indicated that significant baseline risk factors for the time to the first SRE included luminal B type disease (HR = 1.8, P = 0.006), presence of extraskeletal metastasis (HR = 1.7, P = 0.027), prior history of palliative radiation therapy (HR =1.7, P = 0.001), elevation of serum levels of calcium above normal (HR = 1.3, P = 0.002) on administyation of the initial BMA dose, or initiation of BMA treatment above 6 months (HR = 1.4, P = 0.044) after initial detection of bone metastasis. The significance of these factors was also observed in the analyses of multiple SREs. Conclusions: To reduce the risk of SREs, BMAs should be initiated within 6 months of the diagnosis of bone metastasis, and before commencing palliative radiation therapy in breast cancer patients. For patients with luminal B breast cancer and existing extraskeletal metastasis, BMAs should be continued whenever possible. Disclosure: All authors have declared no conflicts of interest.

Abstract P5-08-12: Histopathological and immunohistochemical findings for epithelial-to-mesenchymal transition were associated with clinical progressive disease of triple-negative breast cancers during neoadjuvant chemotherapy

Background : Clinical progressive disease (cPD) occurs during neoadjuvant therapy (NAC) in 3 to 5% of breast cancer patients. By gene expression profiling analyses using DNA microarray, the expression of epithelial-to-mesenchymal transition (EMT)-associated genes were shown to be correlated with chemoresistant phenotype of breast cancer cell lines. In order to reveal clinical implication of EMT-associated molecules on the acquisition of cPD property, we designed a retrospective histopathilogical and immunohistochemical case-control study. Patients and Methods : From pathology database of patients who received surgical resection, 86 patients with early triple-negative breast cancer (TNBC) were identified: 23 patients suffered cPD during NAC (PD group) and 63 control group patients did not receive NAC (C group), in whom >95% of patients was estimated to be non-PD group if NAC was performed. For these 86 cases, histopathological classification was performed and negativity of hormone receptors and HER2 was confirmed. As EMT markers, we immunohistochemically examined the expressions of vimentin, Twist, Twist NB and Snail. The chi-square test was used to assess statistically significant differences between the groups. Results : Histologically, PD group comprised 12 invasive ductal carcinomas (IDCs) (52%) and 11 metaplastic carcinomas (MPCs) (48%), and they all were nuclear grade 3. In C group, 52 (83%) were IDCs and 10 (16%) were histological types other than IDC or MPC, and there was only 1 MPC (1%) ( p = 4.31E-08). Nuclear grade was 1, 2, and 3 in 1, 9, and 53, respectively. Vimentin was positive in cytoplasm of 74% of PD group and the incidence was higher than that in C group (49%) ( p = 0.016). Cytoplasmic twist-2 and cytoplasmic and nuclear Snail expressions were detected almost equally between PD group and C group. Cytoplasmic Twist NB expression was more frequent in PD group (26%) than in C group (1.6%) ( p = 0.0002). In PD group, a total of 17 cases comprising 9 MPCs and 8 IDCs, were positive for vimentin. In these 17 vimentin-positive cases, 14 were stable disease or partial response with an anthracyclin-containing regimen while all 16 patients receiving a taxane-containing regimen became cPD during the taxane-containing regimen. View this table: Patient characteristics Conclusion : EMT features detected by metaplastic phenotype and cytoplasmic vimentin expression could be predictors for cPD during NAC for TNBC. The tumors of these phenotypes were likely to be resistant to a taxane-containing regimen. View this table: Immunohistochemical profiling Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-08-12.