Therapeutic Efficacy of GC1118, a Novel Anti-EGFR Antibody, against Glioblastoma with High EGFR Amplification in Patient-Derived Xenografts.

We aimed to evaluate the preclinical efficacy of GC1118, a novel anti-epidermal growth factor receptor (EGFR) monoclonal antibody (mAb), against glioblastoma (GBM) tumors using patient-derived xenograft (PDX) models. A total of 15 distinct GBM PDX models were used to evaluate the therapeutic efficacy of GC1118. Genomic data derived from PDX models were analyzed to identify potential biomarkers associated with the anti-tumor efficacy of GC1118. A patient-derived cell-based high-throughput drug screening assay was performed to further validate the efficacy of GC1118. Compared to cetuximab, GC1118 exerted comparable growth inhibitory effects on the GBM tumors in the PDX models. We confirmed that GC1118 accumulated within the tumor by crossing the blood-brain barrier in in vivo specimens and observed the survival benefit in GC1118-treated intracranial models. Genomic analysis revealed high EGFR amplification as a potent biomarker for predicting the therapeutic efficacy of GC1118 in GBM tumors. In summary, GC1118 exerted a potent anti-tumor effect on GBM tumors in PDX models, and its therapeutic efficacy was especially pronounced in the tumors with high EGFR amplification. Our study supports the importance of patient stratification based on EGFR copy number variation in clinical trials for GBM. The superiority of GC1118 over other EGFR mAbs in GBM tumors should be assessed in future studies.

Promising Therapeutic Efficacy of GC1118, an Anti-EGFR Antibody, against KRAS Mutation-Driven Colorectal Cancer Patient-Derived Xenografts.

Epidermal growth factor receptor (EGFR)-targeted monoclonal antibodies, including cetuximab and panitumumab, are used to treat metastatic colorectal cancer (mCRC). However, this treatment is only effective for a small subset of mCRC patients positive for the wild-type KRAS GTPase. GC1118 is a novel, fully humanized anti-EGFR IgG1 antibody that displays potent inhibitory effects on high-affinity EGFR ligand-induced signaling and enhanced antibody-mediated cytotoxicity. In this study, using 51 CRC patient-derived xenografts (PDXs), we showed that KRAS mutants expressed remarkably elevated autocrine levels of high-affinity EGFR ligands compared with wild-type KRAS. In three KRAS-mutant CRCPDXs, GC1118 was more effective than cetuximab, whereas the two agents demonstrated comparable efficacy against three wild-type KRAS PDXs. Persistent phosphatidylinositol-3-kinase (PI3K)/AKT signaling was thought to underlie resistance to GC1118. In support of these findings, a preliminary improved anti-cancer response was observed in a CRC PDX harboring mutated KRAS with intrinsically high AKT activity using GC1118 combined with the dual PI3K/mammalian target of rapamycin (mTOR)/AKT inhibitor BEZ-235, without observed toxicity. Taken together, the superior antitumor efficacy of GC1118 alone or in combination with PI3K/mTOR/AKT inhibitors shows great therapeutic potential for the treatment of KRAS-mutant mCRC with elevated ratios of high- to low-affinity EGFR ligands and PI3K-AKT pathway activation.

Vitamin E-Conjugated Phosphopeptide Inhibitor of the Polo-Box Domain of Polo-Like Kinase 1.

Polo-like kinase 1 (Plk1) regulates cell cycle and cell proliferation, and is currently considered a potential biomarker in clinical trials for many cancers. A characteristic feature of Plks is their C-terminal polo-box domain (PBD). Pro-Leu-His-Ser-pThr (PLHS[pT])-the phosphopeptide inhibitor of the PBD of Plk1-induces apoptosis in cancer cells. However, because of the low cell membrane-penetration ability of PLHS[pT], new approaches are required to overcome these drawbacks. We therefore developed a vitamin E (VE) conjugate that is biodegradable by intracellular redox enzymes as an anticancer drug-delivery system. To ensure high efficiency of membrane penetration, we synthesized VE-S-S-PLHS[pT]KY (1) by conjugating PLHS[pT] to VE via a disulfide bond. We found that 1 penetrated cancer cell membranes, blocked cancer cell proliferation, and induced apoptosis in cancer cells through cell cycle arrest in the G2/M phase. We synthesized a radiolabeled peptide (124I-1), and the radioligand was evaluated in in vivo tumor uptake using positron emission tomography. This study shows that combination conjugates are an excellent strategy for specifically targeting Plk PBD. These conjugates have a dual function, with possible uses in anticancer therapy and tumor diagnosis.

Pharmacokinetics, Biodistribution, and Toxicity Evaluation of Anti-SEMA3A (F11) in In Vivo Models.

BACKGROUND/AIM: The aim of our study was to investigate the pharmacokinetics (PK), tissue distribution and toxicity of F11 antibody to semaphorin 3A in mouse models and explore its anti-angiogenic and tumor-inhibitory effect. MATERIALS AND METHODS: Patient-derived xenograft (PDX) models were established via subcutaneous implantation of glioblastoma multiforme (GBM) cells and treated with F11. RESULTS: F11 significantly attenuated tumor growth and angiogenesis in the GBM PDX model. Within the range of administered doses, the PK of F11 in serum demonstrated a linear fashion, consistent with general PK profiles of soluble antigen-targeting antibodies. Additionally, the clearance level was detected at between 4.63 and 7.12 ml/d/kg, while the biological half-life was measured at 6.9 and 9.4 days. Tissue distribution of F11 in kidney, liver and heart was consistent with previously reported antibody patterns. However, the presence of F11 in the brain was an interesting finding. CONCLUSION: Collectively, our results revealed angiogenic and tumor-inhibitory effect of F11 antibody and its potential therapeutic use within a clinical framework based on PK, biodistribution and toxicity evaluation in mouse models.

Antitumor activity, pharmacokinetics, tumor-homing effect, and hepatotoxicity of a species cross-reactive c-Met antibody.

The receptor tyrosine kinase c-Met plays critical roles in promoting tumor growth, invasion, metastasis, and angiogenesis in various types of cancer and is a promising therapeutic target. The development of a species cross-reactive therapeutic antibody could provide useful to comprehensive preclinical assessment in animal models. Towards this goal, we developed human/mouse cross-reactive c-Met antibodies using an antibody phage library. IRCR201, a c-Met antibody with species cross-reactivity, successfully inhibited the HGF/c-Met signaling pathway via degradation of c-Met and disruption of the binding with its partners, and demonstrated strong in vivo antitumor activity. In pharmacokinetic analysis, IRCR201 exhibited a nonlinear pharmacokinetic profile and showed rapid serum clearance at low dosage. Ex vivo fluorescence imaging and immunohistochemistry demonstrated strong tumor accumulation of IRCR201. Hepatotoxicity analysis revealed that IRCR201 does not significantly affect primary human and mouse hepatocytes. Serum chemistry analysis demonstrated that the alanine aminotransferase serum level was elevated in mice treated with 30 mg/kg IRCR201 than in PBS-treated mice, whereas the levels of aspartate aminotransferase and blood urea nitrogen did not significantly differ. Thus, IRCR201 is a potent therapeutic antibody that can disrupt the HGF/c-Met signaling axis and its species cross-reactivity would enable to evaluate precise biological activity in animal models.

Allergen-Removed Rhus verniciflua Extract Induces Ovarian Cancer Cell Death via JNK Activation.

Nuclear factor-[Formula: see text]B (NF-[Formula: see text]B)/Rel transcription factors are best known for their central roles in promoting cell survival in cancer. NF-[Formula: see text]B antagonizes tumor necrosis factor (TNF)-[Formula: see text]-induced apoptosis through a process involving attenuation of the c-Jun-N-terminal kinase (JNK). However, the role of JNK activation in apoptosis induced by negative regulation of NF-[Formula: see text]B is not completely understood. We found that allergen-removed Rhus verniciflua Stokes (aRVS) extract-mediated NF-[Formula: see text]B inhibition induces apoptosis in SKOV-3 ovarian cancer cells via the serial activation of caspases and SKOV-3 cells are most specifically suppressed by aRVS. Here, we show that in addition to activating caspases, aRVS extract negatively modulates the TNF-[Formula: see text]-mediated I[Formula: see text]B/NF-[Formula: see text]B pathway to promote JNK activation, which results in apoptosis. When the cytokine TNF-[Formula: see text] binds to the TNF receptor, I[Formula: see text]B dissociates from NF-[Formula: see text]B. As a result, the active NF-[Formula: see text]B translocates to the nucleus. aRVS extract (0.5[Formula: see text]mg/ml) clearly prevented NF-[Formula: see text]B from mobilizing to the nucleus, resulting in the upregulation of JNK phosphorylation. This subsequently increased Bax activation, leading to marked aRVS-induced apoptosis, whereas the JNK inhibitor SP600125 in aRVS extract treated SKOV-3 cells strongly inhibited Bax. Bax subfamily proteins induced apoptosis through caspase-3. Thus, these results indicate that aRVS extract contains components that inhibit NF-[Formula: see text]B signaling to upregulate JNK activation in ovarian cancer cells and support the potential of aRVS as a therapeutic agent for ovarian cancer.

Anti-miR delivery strategies to bypass the blood-brain barrier in glioblastoma therapy.

Small non-coding RNAs called miRNAs are key regulators in various biological processes, including tumor initiation, propagation, and metastasis in glioblastoma as well as other cancers. Recent studies have shown the potential for oncogenic miRNAs as therapeutic targets in glioblastoma. However, the application of antisense oligomers, or anti-miRs, to the brain is limited due to the blood-brain barrier (BBB), when administered in the traditional systemic manner. To induce a therapeutic effect in glioblastoma, anti-miR therapy requires a robust and effective delivery system to overcome this obstacle. To bypass the BBB, different delivery administration methods for anti-miRs were evaluated. Stereotaxic surgery was performed to administer anti-Let-7 through intratumoral (ITu), intrathecal (ITh), and intraventricular (ICV) routes, and each method's efficacy was determined by changes in the expression of anti-Let-7 target genes as well as by immunohistochemical analysis. ITu administration of anti-miRs led to a high rate of anti-miR delivery to tumors in the brain by both bolus and continuous administration. In addition, ICV administration, compared with ITu administration, showed a greater distribution of the miR across entire brain tissues. This study suggests that local administration methods are a promising strategy for anti-miR treatment and may overcome current limitations in the treatment of glioblastoma in preclinical animal models.

The diagnostic values of ultrasound and ultrasound-guided fine needle aspiration in subcentimeter-sized thyroid nodules.

BACKGROUND: The diagnostic performances of ultrasound (US) examination and US-guided fine needle aspiration (US-FNA) were investigated in thyroid nodules ≤10 mm in size. METHODS: From April 2006 to December 2006, 1440 nodules ≤10 mm in size in 1403 patients (mean age, 49.3 years; range, 10-84 years) underwent US and US-FNA. The association between nodule size and inadequate specimen was investigated using the Cochran-Armitage trend test and multivariate logistic regression analysis. To evaluate the diagnostic performances of US and US-FNA, we selected 852 nodules that had undergone surgery, follow-up US-FNA, or follow-up US. A receiver operating characteristic (ROC) curve analysis was used to determine the accuracies of US and US-FNA. RESULTS: Of 1440 nodules, 256 (17.8%) yielded inadequate specimens. As the nodule size increased, the rate of inadequate specimens decreased (P < 0.001). A size of 6 mm demonstrated statistical significance between inadequate and adequate specimens (P < 0.001). The diagnostic accuracy of US was slightly improved as nodule size increased. The false positive rate of US examination was higher in nodules ≤6 mm compared with that of nodules >6 mm in size (P = 0.049). However, US-FNA demonstrated high diagnostic performance in all nodules with adequate specimen. CONCLUSIONS: The inadequate specimens of US-FNA and false positives for US examinations increased as nodule size decreased. However, US-FNA demonstrated good diagnostic accuracy, assuming the specimen was adequate.

Sonographic features of traumatic neuromas after neck dissection.

PURPOSE: To evaluate the sonographic features of traumatic neuromas after neck dissection. METHODS: This study included 8 patients whose ages ranged from 36-69 years (mean, 49 years). In all cases, traumatic neuromas were incidentally detected at neck sonography for evaluation of suspected recurrence of well-differentiated papillary carcinoma of the thyroid. All sonograms and medical records were retrospectively reviewed. RESULTS: This study covered 8 cases in which traumatic neuromas were diagnosed by clinical, laboratory, fine-needle aspiration biopsy (FNAB), and other imaging modalities. None of the patients had clinical signs of neuromas, which were, incidentally, discovered by neck sonography. A noticeable sonographic feature in all cases was an isoechoic mass with internal parallel heterogeneous hyperechogenicity. All patients complained of severe pain during FNAB. The cytological results of 2 patients showed fragments of nerve tissue. The remaining 6 FNABs were nondiagnostic. Thyroglobulin (Tg) levels in washout fluids from FNAB of all patients were <0.2 ng/mL, indicating nonthyroidal origin. CONCLUSION: Distinctive sonographic features, sharp pain during FNAB, and low Tg levels in FNAB washout fluid can help to diagnose traumatic neuromas without surgery.

Power Doppler sonography: evaluation of solid breast lesions and correlation with lymph node metastasis.

OBJECTIVE: The purpose of this study was to assess the role of power Doppler US in differentiating benign from malignant solid breast masses when used in conjunction with grayscale sonography (US) and the relationship with axillary lymph node metastasis. SUBJECTS AND METHODS: Grayscale US of 353 solid lesions was categorized using the US BI-RADS final assessment system prospectively. On power Doppler US, the presence of identified or penetrating vessels within the mass was evaluated, respectively. Diagnostic accuracy was calculated at grayscale US with and without power Doppler US. Among malignant cases, tumor vascularity was correlated with lymph node involvement. RESULTS: When the size of the masses was controlled, the identified or penetrating vessels were significantly more frequent in malignant lesions (P<.05). By using the identified and penetrating vessels in the mass as one of the diagnostic criteria for malignancy, the diagnostic accuracy was improved. In 54 infiltrating ductal carcinomas, although lymph node involvement was more frequently seen in the group having identified or penetrating vessels (40 and 35.3%) than in the avascular group (16.7 and 27%), it was not statistically significant (P>.05). CONCLUSION: We suggest that identified or penetrating vessels within the mass on power Doppler US can be one of the malignant criteria.

Sonographic detection of thyroid cancer in breast cancer patients.

The purpose of our study was to analyze the incidence of incidental thyroid cancers which were detected by simultaneous sonographic examination of breast and thyroid glands. Between January 2001 and March 2004, 518 patients were diagnosed with breast cancer after modified radical mastectomy (n=369) or breast conserving surgery (n=149). We screened thyroid glands when we examined breast for diagnosis and follow-up after surgery. If we found the sonographic finding of suspicious for malignancy in thyroid, we immediately performed ultrasound-guided fine needle aspiration biopsy (FNAB). Forty-two cases showed suspicious sonographic findings and of those, 18 cases (42.9%) were determined to have suspicious malignant cytology by ultrasound guided FNAB. Among 518 breast cancers, total 13 cases (2.5%) were diagnosed with papillary carcinoma after thyroidectomy. The mean longest diameter of the thyroid masses was 9.9 mm (range 1-30 mm). Six cases (6/13, 46.2%) were diagnosed as simultaneous breast and thyroid cancers, and the rest of the thyroid cancers were detected after 6 to 33 months (mean 16.5 months) after surgery. In conclusion, the patients with breast cancer had a high incidence (2.5%) of thyroid cancer. Sonographic screening is useful for the early detection of thyroid cancer.

Malignant fibrous histiocytoma (MFH) in axilla.

Palpable axilla mass in woman is relatively rare. Almost all palpable lumps in axilla are axillarys accessory breasts without mass lesion. All diseases develop in breast can also develop in axillarys accessory breasts and other soft tissue mass can occur in axilla. Malignant fibrous histiocytoma (MFH) is the most common malignant soft tissue tumor, but axillarys MFH is extremely rare. We report our experience with a 75-year-old woman with MFH in axilla, treated with wide excision.