Hiroaki Senju

Relationship between UGT1A1*27 and UGT1A1*7 polymorphisms and irinotecan-related toxicities in patients with lung cancer

The objective of this study was to evaluate the effects of gene polymorphisms, including UGT1A1*7, *27, and *29, on the safety of irinotecan therapy.

Phase I study of pemetrexed and concurrent radiotherapy for previously untreated elderly patients with locally advanced non-squamous non-small cell lung cancer

Chemoradiotherapy is the standard treatment for locally advanced non‐small cell lung cancer (NSCLC); however, it is disputed whether this treatment is suitable for patients aged ≥75. This study was conducted to determine the maximum tolerated dose (MTD) of pemetrexed for use in concurrent radiotherapy for elderly patients with locally advanced non‐squamous NSCLC.

Live Cell Labeling with Terpyridine Derivative Proligands to Measure Cytotoxicity Mediated by Immune Cells

Immunotherapy using immune checkpoint inhibitors and CAR‐T cells has revolutionized treatment for patients with malignant tumors. However, measuring tumor cell cytotoxicity mediated by immune effector cells in clinical laboratories has been difficult due to the requirement for radioactive substances. In this study, a series of novel terpyridine derivative proligands were synthesized, and a non‐radioactive cellular cytotoxicity assay using the newly synthesized compounds was developed for use in preclinical and clinical studies for cancer immunotherapy. Once internalized into target cells, the compounds are hydrolyzed by esterases, resulting in the intracellular accumulation of the negatively charged terpyridine derivatives. When the labeled target cells are recognized and killed by immune effector cells, the integrity of the cell membrane is disrupted, and the terpyridine derivatives are released. Upon combining the culture supernatant with europium (Eu3+), the cytotoxicity of immune effector cells for the target cells can be quantitatively determined by measuring the intensity of the Eu3+/ligand‐derived time‐resolved fluorescence. Thus, the assay developed in this study would facilitate the development of novel cancer immunotherapies.

Diffuse alveolar hemorrhage with pseudoprogression during nivolumab therapy in a patient with malignant melanoma: Alveolar hemorrhage in nivolumab therapy

Nivolumab, an anti‐PD‐1 antibody, has been shown to be effective in many cancers, such as malignant melanoma and lung cancer; however, nivolumab therapy can result in pseudoprogression. Diffuse alveolar hemorrhage (DAH) is persistent or recurrent pulmonary hemorrhage as a result of drugs, autoimmune diseases, or infections. DAH with pseudoprogression during nivolumab administration has rarely been reported. Herein, we describe our experience with one such case. A 41‐year‐old woman exhibited bloody sputum and ground glass opacities in the lungs along with tumor growth during nivolumab therapy for multiple lung metastases of malignant melanoma. We diagnosed DAH with pseudoprogression as a result of nivolumab and administered steroid therapy. The DAH subsequently improved and the tumor shrank. This case illustrates that nivolumab can cause DAH with pseudoprogression, which can be controlled by steroid therapy. Thus, if bloody sputum and ground glass opacities in the lungs are observed with tumor growth during nivolumab administration, steroid therapy should be considered to control DAH with pseudoprogression.

A case series of histoplasmosis patients with elevated serum soluble interleukin-2 receptor levels

Histoplasmosis is a common endemic mycosis that is usually asymptomatic but occasionally results in severe illness. Histoplasmosis and its causative agent, Histoplasma capsulatum, are found worldwide but rarely in Japan. In recent years, however, the number of histoplasmosis patients in Japan has increased. In addition, to our knowledge, there are no previous reports of increased serum soluble interleukin-2 receptor (sIL-2R) levels in patients with histoplasmosis. We report a case series of histoplasmosis in three Japanese temporary workers in Manzanillo, Mexico. All three patients developed a persistent high fever and general fatigue. Laboratory tests showed increased C-reactive protein levels and mild liver dysfunction. All patients also showed increased soluble interleukin-2 receptor (sIL-2R) levels. Chest computed tomography revealed multiple nodules in both lung fields. All patients were positive for serum anti-Histoplasma antibodies, and two patients were positive for Histoplasma on polymerase chain reaction tests. After treatment that included antifungals, their conditions gradually improved and laboratory data normalized. Although one patient developed respiratory failure, this patient recovered with antifungal therapy in combination with methylprednisolone. Serum sIL-2R levels in all patients gradually declined to normal levels, indicating their recovery from Histoplasma infection. From our experience with these patients, sIL-2R levels may be a useful biomarker for patients with histoplasmosis.

Abstract 2110: Comparing detection of EGFR mutations in lung cancer patients

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC INTRODUCTION: Epidermal growth factor receptor (EGFR) mutations are a strong determinant factor of response to EGFR tyrosine kinase inhibitors in non-small cell lung cancer (NSCLC). Now some methods have been used to detect EGFR mutations and they have approximately the same sensitivity and specificity. In a case, we experienced unmatched results from two methods of detecting EGFR mutations. In this study, we compared data from three methods of detecting mutations. METHODS: We obtained 30 samples of operation, transbronchial lung biopsy, bronchoscopic brushing, pleural effusion and lymph nodes from NSCLC patients. EGFR mutation status was determined by two or three methods (mutant-enriched PCR method, PNA-LNA PCR Clamp method and PCR invader method). RESULTS:m.e.: mutant-enriched PCR, Clamp: PNA-LNA PCR Clamp, invader: PCR invader View this table: EGFR mutations were detected in 12 samples. Mutation statuses of two or three methods were matched in 9 samples, but 3 samples were unmatched. CONCLUSIONS: EGFR mutation statuses were unmatched in same samples. EGFR gene status is essential information for NSCLC patients. We need to take false negative into consideration in the clinical setting and need father deliberation about detecting methods of EGFR mutations. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2110.

Abstract 2124: The analysis of EGFR genetic heterogeneity on mixed adenocarcinoma of the lung

Background: The epidermal growth factor receptor thyrosine kinase inhibitors (EGFR-TKIs) are effective for non-small cell lung cancer (NSCLC) patients harboring sensitive EGFR mutations. However, every patient eventually relapse even if the treatment of EGFR-TKIs was effective remarkably. It have been reported a secondary T790M mutation, MET amplification and some mechanisms attribute to the drug resistance. Intratumor EGFR genetic heterogeneity has also been considered as a potential cause of the acquired resistance. We previously reported a patient with mixed adenocarcinoma of the lung that had different EGFR mutations in papillary, acinar and BAC subtypes. In this study, we analyzed EGFR mutations in each histological subtypes of mixed adenocarcinoma after surgery and pre EGFR-TKIs treatment. Material and Methods: We recruited twelve mixed adenocarcinoma patients who were treated surgical therapy. Tumor cells in different histological subtypes were microdissected from paraffin-embedded surgical specimens with AS LMD system. Genomic DNA was extracted from each sampled area. EGFR mutations, limited to exon19 deletions and L858R mutations, were examined using the simple polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) methods. Results: Acinar, BAC and papillary sanmples were microdissected from six, ten and eleven tumor tissues. Exon19 deletions and L858R mutation were detected in five and one tissues. From acinar, BAC and papillary samples, one exon19 deletion, two exon19 deletions and three exon19 delitions and one L858R were detected. Out of the six mixed adenocarcinoma, five tissues consisted both mutated and non-mutated cells. Conclusions: Our results indicates that 41% of lung adenocarcinoma with mixed subtypes have intratoumor genetic heterogeneity. EGFR mutations were detected most frequently in papillary samples. There may be some relations between histological subtype and EGFR mutation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2124.

Abstract 1800: Analysis of EGFR mutations before and after EGFR-TKIs treatment

Background: Gefitinib is the epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) that has the dramatically effects in selective patients with non-small cell lung cancer (NSCLC). EGFR mutations predict response and survival in patients treated with gefitinib. However, almost of these patients have a relapse within a year after the start of therapy. Although the mechanisms of acquired resistance to Gefitinib were MET amplification, L861Q and hepatocyte growth factor(HGF), main reason was a secondary EGFR mutation, threonine to methionine at codon 790 in exon 20 (T790M). Therefore, we analyzed EGFR mutations after gefitinib treatment in addition to before treatment in patients with NSCLC. Materials and methods: From December 2004 to September 2009, 37 patients with advanced NSCLC harboring EGFR mutations were treated with gefitinib in our hospital. When tumor was relapsed, we got written informed consent from patients, and re-examination of EGFR mutations was performed in 9 patients by present. EGFR mutations were examined by mutant-enriched polymerase chain reaction (PCR) method and/or peptide nucleic acid-locked nucleic acid (PNA-LNA) PCR clamp method. Samples were the paraffin-embedded specimens, transbronchial biopsied tissues, surgically resected tissues, bronchial washing liquids, and pleural effusion, and we used relapsed tumor samples at secondary analysis. In addition, we evaluated the response to treatment according to the Response Evaluation Criteria in Solid Tumors (RECIST) and progression free survival defined as the time from the date of beginning treatment to the date of disease progression or death. Results: 37 patients treated by gefitinib, response rate was 60%(22/37) and disease control rate was 78%(29/37). Within a year, most patients were relapse. In 9 patients, we could analyze EGFR mutations again. Secondary analysis of EGFR mutations was performed in safety. T790M, a resistant mutation of EGFR, was detected in 4 patients. Samples from lymph node biopsy were used in 2 patients, pleural effusion was in a patient, and transbronchial biopsied tissue was in a patient. 1 patient was adenosquamous cell carcinoma, and all of the rest were adenocarcinoma. Despite emergence of T790M, in a patient, readministration of gefitinib was effective (stable disease), furthermore, treatment of erlotinib after gefitinib was also effective (partial response). In 1 patient, secondary analysis is performing at present. Conclusion: These results suggested the importance of EGFR analysis after relapse in patients treated with gefitinib. We must consider why erlotinib was effective in patients with T790M, and screen appropriate subjects of readministration of EGFR-TKIs. The new therapeutic strategy according to the mechanism of resistance to EGFR-TKI is expected. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1800.