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  • 1
    In: Nature Communications, Springer Science and Business Media LLC, Vol. 11, No. 1 ( 2020-01-03)
    Abstract: Despite the promising clinical efficacy of the second-generation anaplastic lymphoma kinase (ALK) inhibitor alectinib in patients with ALK-rearranged lung cancer, some tumor cells survive and eventually relapse, which may be an obstacle to achieving a cure. Limited information is currently available on the mechanisms underlying the initial survival of tumor cells against alectinib. Using patient-derived cell line models, we herein demonstrate that cancer cells survive a treatment with alectinib by activating Yes-associated protein 1 (YAP1), which mediates the expression of the anti-apoptosis factors Mcl-1 and Bcl-xL, and combinatorial inhibition against both YAP1 and ALK provides a longer tumor remission in ALK-rearranged xenografts when compared with alectinib monotherapy. These results suggest that the inhibition of YAP1 is a candidate for combinatorial therapy with ALK inhibitors to achieve complete remission in patients with ALK-rearranged lung cancer.
    Type of Medium: Online Resource
    ISSN: 2041-1723
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2020
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  • 2
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 80, No. 16_Supplement ( 2020-08-15), p. 2728-2728
    Abstract: The brain metastasis of lung cancer is a crucial problem that causes poor clinical outcome and alters patient's quality of life. Previous reports have indicated that tumor formation is promoted by increased expression of “Don't eat me” signals in cancer cells that inhibit phagocytic action of macrophages. Inhibiting the signals and promoting phagocytosis, such as anti-CD47 or anti-CD24a neutralize antibody, have been focused as a novel therapeutic method, but whether the signals can regulate generation of brain metastasis is still unclear. Here, we have developed an in-vivo imaging method that can allow us to follow the tumor fate in single cell resolution to evaluate microglial phagocytosis of tumor. Using this method, we classified various fates of cancer cells invaded into the brain which is partially determined by microglial phagocytotic functions as a therapeutic target. An mCherry-labeled lung cancer cell line (CMT167mC) was established from CMT167, a mice lung cancer cell line derived from C57BL. A craniotomy was performed on CX3CR1-EGFP mice, in which EGFP is specifically expressed in microglia. The skull of the mice was replaced with a glass coverslip for chronic imaging, and the fluorescently labeled cancer cells were injected via the internal carotid artery of the mice to visualize simultaneously with microglia using two-photon microscopy. To detect a microglial phagocytosis of cancer cells in micro-metastasis region and to follow the tumor fate after phagocytosis, we first evaluate the natural fate of the cancer cells after injection into the brain. Arrested cancer cells in the brain vascular were identified immediately after the cancer cell injection. The fate of each arrested cell could be followed for 14 days. Monitoring of 105 arrested cancer cells (N=3 mice) in the brain revealed 54% of cancer cells displaced from their original site, development of micro-metastasis and attached with microglia in 34.3% of cells, and phagocytosis of 13.3% of cells by microglia resulting in cell death, suggesting their heterogeneous fate. To focus on the microglial phagocytosis of cancer cells that may be one of the candidate for therapeutic target, we evaluated the functional role of a “Don't eat me” signal, which is a surface protein CD47 that inhibited the phagocytic action of macrophages when the cancer cells were invaded to the brain. A CD47 gene knockout cell line (CMT167mC-CD47-/-) was established from CMT167mC cells using CRISPR/Cas9. CMT167mC-CD47-/- cells provided into the brain developed micro-metastasizes less frequently as compared with CMT167mC. Further study to address the role of CD47 in microglial phagocytosis is in progress. The result suggests that this system potentially provide a novel screening method for validation of crucial target inhibiting “Don't eat me” signal. Targeting the microglial phagocytosis could be a potent novel therapy to prevent the development of brain metastasis. In future, other potential “Don't eat me” signals, such as CD24a and PD-L1, will be evaluated. Citation Format: Takahiro Tsuji, Hiroaki Wake, Mariko Shindo, Koichiro Haruwaka, Hitomi Ajimizu, Masatoshi Yamazoe, Tomoko Funazo, Yuto Yasuda, Hironori Yoshida, Yuichi Sakamori, Young Hak Kim, Hiroaki Ozasa, Toyohiro Hirai. Novel in vivo imaging method to evaluate “Don't eat me” signal of tumor against microglia [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2728.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2020
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  • 3
    In: Molecular and Clinical Oncology, Spandidos Publications, ( 2017-06-29)
    Type of Medium: Online Resource
    ISSN: 2049-9450 , 2049-9469
    Language: Unknown
    Publisher: Spandidos Publications
    Publication Date: 2017
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  • 4
    In: Cancer Science, Wiley, Vol. 114, No. 2 ( 2023-02), p. 546-560
    Abstract: Tyrosine kinase inhibitors (TKIs) that target the ROS proto‐oncogene 1, receptor tyrosine kinase ( ROS1 ) gene have shown dramatic therapeutic effects in patients with ROS1‐rearranged non‐small‐cell lung cancer (NSCLC). Nevertheless, advanced ROS1‐rearranged NSCLC is rarely cured as a portion of the tumor cells can survive the initial stages of ROS1‐TKI treatment, even after maximum tumor shrinkage. Therefore, understanding the mechanisms underlying initial cell survival during ROS1‐TKI treatment is necessary to prevent cell survival and achieve a cure for ROS1‐rearranged NSCLC. In this study, we clarified the initial survival mechanisms during treatment with lorlatinib, a ROS1 TKI. First, we established a patient‐derived ezrin gene‐ROS1‐rearranged NSCLC cell line (KTOR71). Then, following proteomic analysis, we focused on yes‐associated protein 1 (YAP1), which is a major mediator of the Hippo pathway, as a candidate factor involved in cell survival during early lorlatinib treatment. Yes‐associated protein 1 was activated by short‐term lorlatinib treatment both in vitro and in vivo. Genetic inhibition of YAP1 using siRNA, or pharmacological inhibition of YAP1 function by the YAP1‐inhibitor verteporfin, enhanced the sensitivity of KTOR71 cells to lorlatinib. In addition, the prosurvival effect of YAP1 was exerted through the reactivation of AKT. Finally, combined therapy with verteporfin and lorlatinib was found to achieve significantly sustained tumor remission compared with lorlatinib monotherapy in vivo. These results suggest that YAP1 could mediate initial cell resistance to lorlatinib in KTOR71 cells. Thus, combined therapy targeting both YAP1 and ROS1 could potentially improve the outcome of ROS1‐rearranged NSCLC.
    Type of Medium: Online Resource
    ISSN: 1347-9032 , 1349-7006
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2023
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  • 5
    In: Annals of Oncology, Elsevier BV, Vol. 30 ( 2019-10), p. vi120-
    Type of Medium: Online Resource
    ISSN: 0923-7534
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2019
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  • 6
    Online Resource
    Online Resource
    American Association for Cancer Research (AACR) ; 2018
    In:  Cancer Research Vol. 78, No. 13_Supplement ( 2018-07-01), p. 295-295
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 78, No. 13_Supplement ( 2018-07-01), p. 295-295
    Abstract: Lung cancer is the leading cause of cancer death. Small cell lung cancer (SCLC) is a histologic subtype of lung cancer and the proportion of SCLC is approximately 15%. New molecular targeted drugs and immune checkpoint inhibitors are successfully effective for non-small cell lung cancer. However, treatment of SCLC has not been improved over recent decades. It is important to explore new treatment strategies of SCLC. MCL1 is a member of the BCL-2 family, which regulates apoptosis. Targeting MCL1 represents a potential breakthrough of cancer treatment. We tested S63845, a MCL1 inhibitor, in four SCLC cell lines (DMS114, DMS53, SW1271, and NCI-H69) and, in addition, one patient derived SCLC cell (KTOR201). S63845 had greater efficacy in two of five SCLC cells (DMS114 and KTOR201). These two SCLC cells had higher expression of MCL1 and lower expression of BCL-XL, which is another member of the BCL-2 family. The other three SCLC cells (DMS53, SW1271 and NCI-H69) were resistant to S63845 and had a higher expression of BCL-XL or lower expression of MCL1. These data suggested that S63845 might be a powerful treatment of SCLC as a new therapeutic strategy. It is possible that the expressions of MCL1 and other members of the BCL-2 family predict the sensitivity of S63845. Citation Format: Yuto Yasuda, Hiroaki Ozasa, Takahiro Tsuji, Takashi Nomizo, Tomoko Funazo, Hironori Yoshida, Yuichi Sakamori, Toyohiro Hirai, Young Hak Kim. Efficacy of MCL1 inhibitor S63845 in small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 295.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2018
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  • 7
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 78, No. 13_Supplement ( 2018-07-01), p. 2604-2604
    Abstract: Treatment of lung cancer is improved recently, especially immune-check point inhibitors prolong progression-free survival (PFS) and overall survival (OS), however, Lung cancer is still leading cause of death worldwide. This is because of response rates in the treatment of non-small cell lung cancer (NSCLC) with combination platinum-based chemotherapy vary from 20% to 40%, and response rates of the treatment of NSCLC with nivolumab is around 20%, whereas with leaving a large number of patients with either stable or progressive disease. It is currently difficult to predict treatment response to chemotherapy and nivolumab since there are no precise biomarkers for it. Recently, we have reported that PD-L1 single nucleotide polymorphisms (SNPs) are associated with response to nivolumab treatment. This study was intended to determine the efficacy of platinum-based combination chemotherapy either in a doublet or triplet, and nivolumab treatment respect to PD-L1 SNPs among patients with NSCLC. A total of 139 patients with NSCLC were treated with platinum-based doublet or triplet chemotherapy, 73 patients treated with nivolumab and were also evaluated for PD-L1 SNPs from plasma DNA. We investigated the association among PD-L1 SNPs, objective response rate (ORR) and PFS. PD-L1 rs2282055 was associated with ORR and PFS in the patients treated with pulatinum-based chemotherapy and nivolumab. In the patients treated with nivolumab, the ORR was 24%, 12%, and 0% for the G/G, G/T and T/T genotypes of PD-L1 rs2282055, respectively. The G allele of PD-L1 rs2282055 was significantly associated with better clinical response compared with the T allele (P = 0.0056 [Cochran-Armitage trend test]). The median PFS time was 2.1 months (95% confidence interval [CI] , 1.8 months to 3.9 months) for the G/G and G/T genotypes and 2.2 months (95% confidence interval [CI], 0.9 months to 2.6 months) for the T/T genotype (P = 0.0210). On the other hand, The T allele of PD-L1 rs2282055 was significantly associated with better clinical response compared with the G allele in the patients treated with platinum-based combination chemotherapy either in a doublet or triplet (P = 0.0080 [Cochran-Armitage trend test] ). The median PFS time was 11.0 months (95% confidence interval [CI], 6.2 months to 16.3 months) for the T/T genotypes and 7.3 months (95% confidence interval [CI] , 6.0 months to 8.2 months) for the G/T and G/G genotype (P = 0.0284). In conclusion, these results suggest that the T/T genotype of PD-L1 SNP rs2282055 associated with the better treatment effect of platinum-based combination chemotherapy, on the contrary, the T/T of rs2282055 negatively associated with response to nivolumab treatment. It might be used as a biomaker for selection of the regimen of NSCLC treatment. Citation Format: Takashi Nomizo, Hiroaki Ozasa, Takahiro Tsuji, Tomoko Funazo, Yuto Yasuda, Hironori Yoshida, Yuichi Sakamori, Toyohiro Hirai, Yong Hak Kim. PD-L1 rs2282055 is associated with opposite treatment effect between platinum-based chemotherapy and nivolumab treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2604.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2018
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  • 8
    In: Oncotarget, Impact Journals, LLC, Vol. 8, No. 42 ( 2017-09-22), p. 71805-71816
    Type of Medium: Online Resource
    ISSN: 1949-2553
    URL: Issue
    Language: English
    Publisher: Impact Journals, LLC
    Publication Date: 2017
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  • 9
    Online Resource
    Online Resource
    Elsevier BV ; 2020
    In:  The Annals of Thoracic Surgery Vol. 110, No. 1 ( 2020-07), p. e39-e41
    In: The Annals of Thoracic Surgery, Elsevier BV, Vol. 110, No. 1 ( 2020-07), p. e39-e41
    Type of Medium: Online Resource
    ISSN: 0003-4975
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    Language: English
    Publisher: Elsevier BV
    Publication Date: 2020
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  • 10
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 82, No. 12_Supplement ( 2022-06-15), p. 1601-1601
    Abstract: Overcoming the treatment resistance of cancer is a problem to be solved in improving the prognosis of cancer patients. Recently, genome analysis has revealed that multiple clusters exist in one tumor, and it has been reported that intratumor heterogeneity causes treatment resistance even in lung cancer. However, the mechanism of intratumor heterogeneity has not yet been clarified. Previous studies of intratumor heterogeneity have mainly focused on analyzing cancer cells from patients, classifying clusters of different properties, and identifying factors that define their characteristics. Multiple clusters are present in the tumor tissue of a single patient, and analysis of multiple patients classifies them into even more clusters, but it is difficult to research all of them. To solve this problem, an early model of intratumor heterogeneity in which homogeneous cells become heterogeneous is needed.We hypothesized that intratumor heterogeneity cell model could be found in resistant strains.Alectinib was exposed in vitro to an ALK-positive lung cancer cell line (H2228) to create an alectinib-resistant cell line (H2228-AR1S). H2228-AR1S has two subpopulations that look different. Of the two subpopulations, the smaller cells have high CD47 expression (CD47 high subpopulation), and the scattered spindle-shaped cells have low CD47 expression (CD47 low subpopulation). Using flow cytometry, each subpopulation was isolated, and its properties were investigated. There was no difference in sensitivity to alectinib between the CD47 high subpopulation and the CD47 low subpopulation. In the low subpopulation of CD47, epithelial markers were decreased, and mesenchymal markers were increased using immunoblotting. It suggests that CD47 low subpopulation has undergone epithelial-mesenchymal transition (EMT). The CD47 high subpopulation had high sphere formation ability in vitro, and high tumorigenicity using Xenograft model. CD47 gene inhibition using siRNA reduced the sphere formation ability of the CD47 high subpopulation. This suggests that CD47 is involved in sphere formation.There have been no reports of CD47 being involved in the characteristics of intratumor heterogeneity. Furthermore, using intratumor heterogeneity cell model, we are exploring the mechanism that regulate division into two subpopulations. Citation Format: Tomoko Y. Funazo, Hiroaki Ozasa, Kentaro Hashimoto, Hiroshi Yoshida, Tatsuya Ogimoto, Kazutaka Hosoya, Hitomi Ajimizu, Takahiro Tsuji, Hironori Yoshida, Ryo Itotani, Yuichi Sakamori, Toyohiro Hirai. CD47 related to intratumor heterogeneity in alectinib-resistant ALK-rearranged lung cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1601.
    Type of Medium: Online Resource
    ISSN: 1538-7445
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2022
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