Noriko Hirai

Interstitial lung disease associated with human papillomavirus vaccination

Vaccinations against the human papillomavirus (HPV) have been recommended for the prevention of cervical cancer. HPV-16/18 AS04-adjuvanted vaccines (Cervarix) are said to have favourable safety profiles. Interstitial lung diseases (ILDs) can occur following exposure to a drug or a biological agent. We report a case of ILD associated with a Cervarix vaccination. A woman in her 40s, with a history of conisation, received three inoculations of Cervarix. Three months later, she presented with a cough and shortness of breath. Findings from a computed tomography of the chest and a transbronchial lung biopsy were consistent with non-specific interstitial pneumonia. Workup eliminated all other causes of the ILD, except for the vaccination. Over the 11 months of the follow-up period, her symptoms resolved without steroid therapy. The onset and spontaneous resolution of the ILD showed a chronological association with the HPV vaccination. The semi-quantitative algorithm revealed that the likelihood of an adverse drug reaction to Cervarix was “Probable”. The outcome was relatively good, but more attention should be paid to a potential risk for HPV vaccinations to cause ILDs. Wherever possible, chest radiographic examinations should be performed in order not to overlook any ILDs.

Abstract 749: NovelALKspecific mRNAin situhybridization assay for non-small cell lung carcinoma

A recent technical advance in the mRNA in situ hybridization (mRNA-ISH) assay provides simultaneous signal amplification and background suppression with a unique probe design to achieve single molecule visualization. We assessed the utility of the mRNA-ISH assay as a diagnostic tool to detect anaplastic lymphoma receptor tyrosine kinase (ALK) mRNA in non-small cell lung carcinoma. We compared the mRNA-ISH assay with immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). The study included 279 surgically resected lung adenocarcinomas and 44 transbronchial biopsied (TBB) adenocarcinomas. mRNA-ISH was conducted using the RNAscope 2.0 system which includes pre-designed detection probes for the tyrosine kinase domain of ALK mRNA. IHC was conducted on all of the 323 samples using ALK-specific antibodies. mRNA-ISH was performed on 279 surgical samples and 6 TBB samples. Break-apart FISH was used to assess cases that were mRNA-ISH or IHC positive. ALK protein expression was detected in 11/279 (3.9%) specimens. ALK mRNA was also detected by mRNA-ISH in these cases, and 9 of the 11 specimens (81%), were also positive by ALK FISH. Using the IHC results as a reference, the sensitivity and specificity of mRNA-ISH was 100%. In the TBB cohort, ALK protein expression was observed in 3/44 (6.8%) of specimens, in which ALK mRNA expression was also detected. ALK mRNA-ISH data were highly correlated with the IHC data, and ALK mRNA-ISH was able to identify every FISH-positive sample. We conclude that mRNA-ISH could play an alternative or complementary role in ALK genetic diagnosis in NSCLC. Citation Format: Noriko Hirai, Takaaki Sasaki, Yoshinobu Ohsaki. Novel ALK specific mRNA in situ hybridization assay for non-small cell lung carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 749. doi:10.1158/1538-7445.AM2017-749

Abstract 4656: The novel mRNA in situ hybridization method for the detection of ALK, RET, ROS1 and NTRK1 mRNA in non-small cell lung cancer

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA [Background]In recent years, tumor genotyping is critical for making decisions of non-small cell lung cancer (NSCLC) treatment. However genetic rearrangement of ALK, RET, ROS1 and NTRK1 were identified in the subset of NSCLC patients, these clinical characteristics were still remained to be clear. Immunohistochemistry (IHC) is widely used as a simple and quick method for screening for protein expression of these oncogenes. Fluorescence in situ hybridization (FISH) or RT-PCR are performed consecutively to confirm the genotyping. However it is often taking time and expensive and it is needed to consolidate novel substitute methods. [Purpose]We performed the in situ hybridization (ISH) analysis to detect cellular mRNA in formalin-fixed paraffin-embedded (FFPE) tissue samples of lung cancer. We also conducted IHC with the use of ALK, RET, ROS1 and NTRK1 specific antibodies and investigate a comparative study to assess the usefulness of mRNA-ISH to detect expression of these mRNA. [Materials and Methods]We made tissue micro array (TMA) slides using 40 FFPE samples of resected lung adenocarcinoma with known epidermal growth factor receptor (EGFR) mutation status by PCR (PNA-LNA PCR clamp method). The mRNA in situ hybridization was employed using RNA scope R 2.0 Regant Kit with targeting probes. IHC staining was conducted by antibodies using 5A4 and D5F3 for ALK, RET01 for RET, D4D6 for ROS1, and 14G6 for NTRK1. [Results]All of the IHC stained cases were clearly detected by mRNA-ISH respectively. There were some cases which were only visualized by ISH, each case was partially positive and not totally of the tumor. ALK: 5% of cases (2 out of 40) were detected in mRNA-ISH and were strongly stained with both 5A4 and D5F3. RET: 20% of cases (8/40) were detected in mRNA-ISH. Two cases (5% of total) had positive RET01 staining, while 6 cases had negative staining. (3 had no other genetic alterations, 2 had EGFR-L858R confirmed by PCR, and 1 had ALK by IHC and FISH.) ROS1: 20% (8/40) were detected in mRNA-ISH. Three cases (7.5% of total) presented positive D4D6 staining, while 5 had negative staining. (3 had no other alterations, 1 had EGFR-E746-A750 and 1 had EGFR-L858R by PCR.) NTRK1: 17.5% of cases (7/40) were detected in mRNA-ISH. 1 case (2.5% of total) presented positive 14G6 staining, while 6 cases had negative staining. (2 had no other alterations, 2 had EGFR-L858R, 1 had EGFR-E746-A750, and 1 had ALK.) [Conclusion] The novel mRNA in situ hybridization method combined with traditional IHC could improve detection rate of ALK, RET, ROS1 and NTRK1 gene alterations in NSCLC FFPE samples. Citation Format: Noriko Hirai, Takaaki Sasaki, Yoshinori Minami, Yoshinobu Ohsaki. The novel mRNA in situ hybridization method for the detection of ALK, RET, ROS1 and NTRK1 mRNA in non-small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4656. doi:10.1158/1538-7445.AM2014-4656