124 results on '"Goi, K"'
Search Results
2. Oncogenic fusion E2A-HLF sensitizes t(17;19)-positive acute lymphoblastic leukemia to TRAIL-mediated apoptosis by upregulating the expression of death receptors
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Zhang, X, Inukai, T, Hirose, K, Akahane, K, Kuroda, I, Honna-Oshiro, H, Kagami, K, Goi, K, Nakamura, K, Kobayashi, M, Endo, M, Yagita, H, Kurosawa, H, Thomas Look, A, Honda, H, Inaba, T, Nakazawa, S, and Sugita, K
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- 2012
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3. Specific induction of CD33 expression by E2A–HLF: the first evidence for aberrant myeloid antigen expression in ALL by a fusion transcription factor
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Akahane, K, Inukai, T, Inaba, T, Kurosawa, H, Look, A T, Kiyokawa, N, Fujimoto, J, Goto, H, Endo, M, Zhang, X, Hirose, K, Kuroda, I, Honna, H, Kagami, K, Goi, K, Nakazawa, S, and Sugita, K
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- 2010
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4. Hypercalcemia in childhood acute lymphoblastic leukemia: frequent implication of parathyroid hormone-related peptide and E2A-HLF from translocation 17;19
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Inukai, T, Hirose, K, Inaba, T, Kurosawa, H, Hama, A, Inada, H, Chin, M, Nagatoshi, Y, Ohtsuka, Y, Oda, M, Goto, H, Endo, M, Morimoto, A, Imaizumi, M, Kawamura, N, Miyajima, Y, Ohtake, M, Miyaji, R, Saito, M, Tawa, A, Yanai, F, Goi, K, Nakazawa, S, and Sugita, K
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- 2007
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5. Resistance of infant leukemia with MLL rearrangement to tumor necrosis factor-related apoptosis-inducing ligand: a possible mechanism for poor sensitivity to antitumor immunity
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Inukai, T, Zhang, X, Goto, M, Hirose, K, Uno, K, Akahane, K, Nemoto, A, Goi, K, Sato, H, Takahashi, K, Honna, H, Kagami, K, Nakamoto, K, Yagita, H, Okumura, K, Koyama-Okazaki, T, Nakazawa, S, and Sugita, K
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- 2006
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6. Association of aberrant ASNS imprinting with asparaginase sensitivity and chromosomal abnormality in childhood BCP-ALL
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Watanabe, A., Miyake, K., Nordlund, Jessica, Syvänen, Ann-Christine, van der Weyden, L., Honda, H., Yamasaki, N., Nagamachi, A., Inaba, T., Ikawa, T., Urayama, K. Y., Kiyokawa, N., Ohara, A., Kimura, S., Kubota, Y., Takita, J., Goto, H., Sakaguchi, K., Minegishi, M., Iwamoto, S., Shinohara, T., Kagami, K., Abe, M., Akahane, K., Goi, K., Sugita, K., Inukai, T., Watanabe, A., Miyake, K., Nordlund, Jessica, Syvänen, Ann-Christine, van der Weyden, L., Honda, H., Yamasaki, N., Nagamachi, A., Inaba, T., Ikawa, T., Urayama, K. Y., Kiyokawa, N., Ohara, A., Kimura, S., Kubota, Y., Takita, J., Goto, H., Sakaguchi, K., Minegishi, M., Iwamoto, S., Shinohara, T., Kagami, K., Abe, M., Akahane, K., Goi, K., Sugita, K., and Inukai, T.
- Abstract
Karyotype is an important prognostic factor in childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL), but the underlying pharmacogenomics remain unknown. Asparaginase is an integral component in current chemotherapy for childhood BCP-ALL. Asparaginase therapy depletes serum asparagine. Normal hematopoietic cells can produce asparagine by asparagine synthetase (ASNS) activity, but ALL cells are unable to synthesize adequate amounts of asparagine. The ASNS gene has a typical CpG island in its promoter. Thus, methylation of the ASNS CpG island could be one of the epigenetic mechanisms for ASNS gene silencing in BCP-ALL. To gain deep insights into the pharmacogenomics of asparaginase therapy, we investigated the association of ASNS methylation status with asparaginase sensitivity. The ASNS CpG island is largely unmethylated in normal hematopoietic cells, but it is allele-specifically methylated in BCP-ALL cells. The ASNS gene is located at 7q21, an evolutionally conserved imprinted gene cluster. ASNS methylation in childhood BCP-ALL is associated with an aberrant methylation of the imprinted gene cluster at 7q21. Aberrant methylation of mouse Asns and a syntenic imprinted gene cluster is also confirmed in leukemic spleen samples from ETV6-RUNX1 knockin mice. In 3 childhood BCP-ALL cohorts, ASNS is highly methylated in BCP-ALL patients with favorable karyotypes but is mostly unmethylated in BCP-ALL patients with poor prognostic karyotypes. Higher ASNS methylation is associated with higher L-asparaginase sensitivity in BCP-ALL through lower ASNS gene and protein expression levels. These observations demonstrate that silencing of the ASNS gene as a result of aberrant imprinting is a pharmacogenetic mechanism for the leukemia-specific activity of asparaginase therapy in BCP-ALL.
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- 2020
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7. The JAK2 inhibitor AG490 predominantly abrogates the growth of human B-precursor leukemic cells with 11q23 translocation or Philadelphia chromosome
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Miyamoto, N, Sugita, K, Goi, K, Inukai, T, Iijima, K, Tezuka, T, Kojika, S, Nakamura, M, Kagami, K, and Nakazawa, S
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- 2001
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8. Abnormalities of the p16INK4a gene in childhood B-precursor acute lymphoblastic leukemia without nonrandom translocations: analysis of seven matched pairs of primary leukemia and corresponding cell line
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Nakamura, M, Sugita, K, Inukai, T, Goi, K, Miyamoto, N, Kagami, K, Okazaki-Koyama, T, Mori, T, Saito, M, and Nakazawa, S
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- 2001
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9. Expression of thrombopoietin receptor and its functional role in human B-precursor leukemia cells with 11q23 translocation or Philadelphia chromosome
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Iijima, K, Sugita, K, Inukai, T, Goi, K, Tezuka, T, Uno, K, Sato, H, Kagami, K, and Nakazawa, S
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- 2000
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10. Participation of granulocyte colony-stimulating factor in the growth regulation of leukemia cells from Philadelphia chromosome-positive acute leukemia and blast crisis of chronic myeloid leukemia
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Inukai, T, Sugita, K, Mitsui, K, Iijima, K, Goi, K, Tezuka, T, Kojika, S, Kagami, K, Mori, T, Kinoshita, A, Suzuki, T, Okazaki-Koyama, T, and Nakazawa, S
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- 2000
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11. p16/MTS1/INK4A gene is frequently inactivated by hypermethylation in childhood acute lymphoblastic leukemia with 11q23 translocation
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Nakamura, M, Sugita, K, Inukai, T, Goi, K, Iijima, K, Tezuka, T, Kojika, S, Shiraishi, K, Miyamoto, N, Karakida, N, Kagami, K, O-Koyama, T, Mori, T, and Nakazawa, S
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- 1999
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12. The KOR-SA3544 antigen predominantly expressed on the surface of Philadelphia chromosome-positive acute lymphoblastic leukemia cells is nonspecific cross-reacting antigen-50/90 (CD66c) and invariably expressed in cytoplasm of human leukemia cells
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Sugita, K, Mori, T, Yokota, S, Kuroki, Ma, O-Koyama, T, Inukai, T, Iijima, K, Goi, K, Tezuka, T, Kojika, S, Shiraishi, K, Nakamura, M, Miyamoto, N, Karakida, N, Kagami, K, and Nakazawa, S
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- 1999
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13. Leukemic cells with 11q23 translocations express granulocyte colony-stimulating factor (G-CSF) receptor and their proliferation is stimulated with G-CSF
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Inukai, T, Sugita, K, Iijima, K, Goi, K, Tezuka, T, Kojika, S, Kagami, K, Mori, T, Kinoshita, A, Suzuki, T, O-Koyama, T, and Nakazawa, S
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- 1998
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14. Sequential glucosylation determined by NMR in the biosynthesis of mulberroside D, a cis-oxyresveratrol diglucoside, in Morus alba L. cell cultures
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Hano, Y., Goi, K., Nomura, T., and Ueda, S.
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- 1997
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15. Clonotypic analysis of acute lymphoblastic leukemia with a double TEL-AML1 fusion at onset and relapse
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Inukai, T, Yokota, S, Okamoto, T, Nemoto, A, Akahane, K, Takahashi, K, Sato, H, Goi, K, Nakazawa, S, and Sugita, K
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- 2006
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16. Microstructure formation of porous sintered Ti-Si-Zr compacts with mechanically alloyed-activated Ti-Si and TiH2 powders
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Jarfors, A. E. W., Butler, D. L., Goi, K. L. S., Jarfors, A. E. W., Butler, D. L., and Goi, K. L. S.
- Abstract
Metallic implants are widely used in applications associated with bone. A major drawback of using metals is their elastic modulus which is higher than that of bone resulting in stress shielding and premature failure of the implant. The employment of biomaterials with a porous structure has the potential to lower the modulus and promote osseointegration. The present work investigates the microstructure formation and the resulting elastic modulus of a new Ti-Si-Zr alloy. The sintering procedure involves the use of both mechanically alloyed Ti-Si powder and TiH2 to activate sintering with the TiH2 also serving as a pore precursor. The procedure is designed to promote bonding but not consolidation. The influence of sintering temperature, heating rate, as well as the amount and size of the TiH2 on the phases formed and porosity was investigated. It was observed that the use of TiH2 increased the degree of porosity whilst the size of TiH2 particles could be used to control the pore size. The results showed that when using small TiH2 particles, the elastic modulus was strongly dependent on the fraction of TiH2. When large TiH2 particles were used, the porosity had no significant influence on the elastic modulus. The variation in behavior could be attributed to differences in microstructure. To control the bulk modulus it is essential to understand the differences in the microstructure formation mechanisms between these two cases. (C) 2014 Elsevier B.V. All rights reserved.
- Published
- 2014
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17. Elastic modulus of sintered porous Ti-Si-Zr, using activation by Ti-Si mechanically alloyed powder and TiH2 powder
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Goi, K. L. S., Butler, D. L., Jarfors, A. E. W., Yong, J. M. S., Lim, D. C. S., Goi, K. L. S., Butler, D. L., Jarfors, A. E. W., Yong, J. M. S., and Lim, D. C. S.
- Abstract
A novel biomaterial based on Ti-Si-Zr was developed using the sintering process with a composition targeting at a bulk modulus in the same range as that of human bone, i.e. 10-30 GPa. Control of porosity should also be possible to allow for the promotion osseointegration. The sintering procedure involves the use of mechanically alloyed Ti-Si-powder, and TiH2, to promote bonding, but not consolidation. The effect of porosity on the bulk modulus using compression testing is investigated. The influence of sintering temperature, heating rate, and amount and size of the TiH2-activator on porosity are also investigated. The achievable bulk modulus was in the range of 20-55 GPa at porosity levels ranging from 16% to 54%. Porosity had a profound influence on the bulk modulus, and the choice of appropriate processing conditions enables the creation of an engineered porosity and bulk modulus primarily by varying the sintering temperature and the size of the TiH2-powder particles. © 2007.
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- 2008
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18. BCR-ABL regulates death receptor expression for TNF-related apoptosis-inducing ligand (TRAIL) in Philadelphia chromosome-positive leukemia
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Kuroda, I, primary, Inukai, T, additional, Zhang, X, additional, Kikuchi, J, additional, Furukawa, Y, additional, Nemoto, A, additional, Akahane, K, additional, Hirose, K, additional, Honna-Oshiro, H, additional, Goi, K, additional, Kagami, K, additional, Yagita, H, additional, Tauchi, T, additional, Maeda, Y, additional, and Sugita, K, additional
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- 2012
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19. Hypercalcemia in childhood acute lymphoblastic leukemia: frequent implication of parathyroid hormone-related peptide and E2A-HLF from translocation 17;19
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Inukai, T, primary, Hirose, K, additional, Inaba, T, additional, Kurosawa, H, additional, Hama, A, additional, Inada, H, additional, Chin, M, additional, Nagatoshi, Y, additional, Ohtsuka, Y, additional, Oda, M, additional, Goto, H, additional, Endo, M, additional, Morimoto, A, additional, Imaizumi, M, additional, Kawamura, N, additional, Miyajima, Y, additional, Ohtake, M, additional, Miyaji, R, additional, Saito, M, additional, Tawa, A, additional, Yanai, F, additional, Goi, K, additional, Nakazawa, S, additional, and Sugita, K, additional
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- 2006
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20. Clonotypic analysis of acute lymphoblastic leukemia with a double TEL-AML1 fusion at onset and relapse
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Inukai, T, primary, Yokota, S, additional, Okamoto, T, additional, Nemoto, A, additional, Akahane, K, additional, Takahashi, K, additional, Sato, H, additional, Goi, K, additional, Nakazawa, S, additional, and Sugita, K, additional
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- 2005
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21. Ligand activation of peroxisome proliferator-activated receptor γ induces apoptosis of leukemia cells by down-regulating the c-myc gene expression via blockade of the Tcf-4 activity
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Yamakawa-Karakida, N, primary, Sugita, K, additional, Inukai, T, additional, Goi, K, additional, Nakamura, M, additional, Uno, K, additional, Sato, H, additional, Kagami, K, additional, Barker, N, additional, and Nakazawa, S, additional
- Published
- 2002
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22. Ligand activation of peroxisome proliferator-activated receptor induces γ apoptosis of leukemia cells by down-regulating the c-myc gene expression via blockade of the Tcf-4 activity.
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Yamakawa-Karakida, N., Sugita, K., Inukai, T., Goi, K., Nakamura, M., Uno, K., Sato, H., Kagami, K., Barker, N., and Nakazawa, S.
- Subjects
APOPTOSIS ,LEUKEMIA ,CANCER cells ,LIGANDS (Biochemistry) - Abstract
The peroxisome proliferator-activated receptor γ (PPAR γ), a member of the nuclear receptor superfamily, is expressed at highest levels in adipose tissue and functions as a central regulator in the process of adipocyte differentiation. In the present study, we showed that human leukemic cell lines, not only myeloid but also lymphoid, express PPAR γ and its activation by natural ligand (15-deoxy-δ[sup 12,14]-prostaglandin J2) and synthetic ligand (troglitazone) profoundly inhibited their proliferation by induction of apoptosis preferentially in the serum-free culture. We pursued its mechanism using the representative cell lines, and found that induction of apoptosis was accompanied by caspase-3 activation and specifically blocked by its inhibitor. While status of several apoptosisrelated molecules remained unchanged, the c-Myc expression was markedly down-regulated within 24 h after troglitazone treatment. The c-myc mRNA levels were dramatically reduced at 1 h and became undetectable at 12 h after troglitazone treatment, which proved to be accompanied by complete blockade of the Tcf-4 activity in the electrophoretic mobility shift assay. We succeeded in establishing HL-60 cell lines growing well in the presence of troglitazone in the long-term serum-free culture. They showed neither induction of apoptosis nor down-regulation of the c-Myc expression via blockade of the Tcf-4 activity after troglitazone treatment. This is the first identification of the linkage between PPAR γ-mediated apoptosis and down-regulation of the c-myc gene expression. [ABSTRACT FROM AUTHOR]
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- 2002
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23. The E2A-HLF oncoprotein activates Groucho-related genes and suppresses Runx1.
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Dang, J, Inukai, T, Kurosawa, H, Goi, K, Inaba, T, Lenny, N T, Downing, J R, Stifani, S, and Look, A T
- Abstract
The E2A-HLF fusion gene, formed by the t(17;19)(q22;p13) chromosomal translocation in leukemic pro-B cells, encodes a chimeric transcription factor consisting of the transactivation domain of E2A linked to the bZIP DNA-binding and protein dimerization domain of hepatic leukemia factor (HLF). This oncoprotein blocks apoptosis induced by growth factor deprivation or irradiation, but the mechanism for this effect remains unclear. We therefore performed representational difference analysis (RDA) to identify downstream genetic targets of E2A-HLF, using a murine FL5.12 pro-B cell line that had been stably transfected with E2A-HLF cDNA under the control of a zinc-regulated metallothionein promoter. Two RDA clones, designated RDA1 and RDA3, were differentially upregulated in E2A-HLF-positive cells after zinc induction. The corresponding cDNAs encoded two WD40 repeat-containing proteins, Grg2 and Grg6. Both are related to the Drosophila protein Groucho, a transcriptional corepressor that lacks DNA-binding activity on its own but can act in concert with other proteins to regulate embryologic development of the fly. Expression of both Grg2 and Grg6 was upregulated 10- to 50-fold by E2A-HLF. Immunoblot analysis detected increased amounts of two additional Groucho-related proteins, Grg1 and Grg4, in cells expressing E2A-HLF. A mutant E2A-HLF protein with a disabled DNA-binding region also mediated pro-B cell survival and activated Groucho-related genes. Among the transcription factors known to interact with Groucho-related protein, only RUNX1 was appreciably downregulated by E2A-HLF. Our results identify a highly conserved family of transcriptional corepressors that are activated by E2A-HLF, and they suggest that downregulation of RUNX1 may contribute to E2A-HLF-mediated leukemogenesis.
- Published
- 2001
24. Acquired copy number amplification at the MYC enhancer in human B-precursor acute lymphoblastic leukemia cell lines.
- Author
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Watanabe A, Wang L, Tan TK, Urayama KY, Kizuki T, Komatsu C, Kagami K, Shinohara T, Kasai S, Tamai M, Harama D, Akahane K, Goi K, Goto H, Satou K, Kaname T, Sanda T, and Inukai T
- Abstract
Our study highlights the discovery of recurrent copy number alterations in noncoding regions, specifically blood enhancer cluster (BENC-CNA), in B-precursor acute lymphoblastic leukemia (BCP-ALL) cell lines. We demonstrate that BENC-CNA acts as a super-enhancer, driving MYC expression and possibly contributing to the immortalization and proliferative advantage of BCP-ALL cells in vitro., (© 2024 The Author(s). Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)
- Published
- 2024
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25. Application of prime editing system to introduce TP53 R248Q hotspot mutation in acute lymphoblastic leukemia cell line.
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Nguyen T, Aida T, Iijima-Yamashita Y, Tamai M, Nagamachi A, Kagami K, Komatsu C, Kasai S, Akahane K, Goi K, Inaba T, Sanada M, and Inukai T
- Subjects
- Humans, Cell Line, Tumor, Drug Resistance, Neoplasm genetics, CRISPR-Cas Systems genetics, RNA, Guide, CRISPR-Cas Systems genetics, Proto-Oncogene Proteins c-mdm2 genetics, Tumor Suppressor Protein p53 genetics, Gene Editing methods, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma pathology, Precursor Cell Lymphoblastic Leukemia-Lymphoma therapy, Mutation
- Abstract
In childhood acute lymphoblastic leukemia (ALL), TP53 gene mutation is associated with chemoresistance in a certain population of relapsed cases. To directly verify the association of TP53 gene mutation with chemoresistance of relapsed childhood ALL cases and improve their prognosis, the development of appropriate human leukemia models having TP53 mutation in the intrinsic gene is required. Here, we sought to introduce R248Q hotspot mutation into the intrinsic TP53 gene in an ALL cell line, 697, by applying a prime editing (PE) system, which is a versatile genome editing technology. The PE2 system uses an artificial fusion of nickase Cas9 and reverse-transcriptase to directly place new genetic information into a target site through a reverse transcriptase template in the prime editing guide RNA (pegRNA). Moreover, in the advanced PE3b system, single guide RNA (sgRNA) matching the edited sequence is also introduced to improve editing efficiency. The initially obtained MDM2 inhibitor-resistant PE3b-transfected subline revealed disrupted p53 transactivation activity, reduced p53 target gene expression, and acquired resistance to chemotherapeutic agents and irradiation. Although the majority of the subline acquired the designed R248Q and adjacent silent mutations, the insertion of the palindromic sequence in the scaffold hairpin structure of pegRNA and the overlap of the original genomic DNA sequence were frequently observed. Targeted next-generation sequencing reconfirmed frequent edit errors in both PE2 and PE3b-transfected 697 cells, and it revealed frequent successful edits in HEK293T cells. These observations suggest a requirement for further modification of the PE2 and PE3b systems for accurate editing in leukemic cells., (© 2024 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)
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- 2024
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26. Involvement of BCR::ABL1 in laminin adhesion of Philadelphia chromosome-positive acute lymphoblastic leukemia through upregulation of integrin α6.
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Takahashi K, Nguyen TTT, Watanabe A, Sato H, Saito K, Tamai M, Harama D, Kasai S, Akahane K, Goi K, Kagami K, Abe M, Komatsu C, Maeda Y, Sugita K, and Inukai T
- Subjects
- Animals, Humans, Mice, Imatinib Mesylate pharmacology, Imatinib Mesylate therapeutic use, Integrin alpha6 genetics, Laminin metabolism, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Up-Regulation
- Abstract
Background: Adhesion of cancer cells to extracellular matrix laminin through the integrin superfamily reportedly induces drug resistance. Heterodimers of integrin α6 (CD49f) with integrin β1 (CD29) or β4 (CD104) are major functional receptors for laminin. Higher CD49f expression is reportedly associated with a poorer response to induction therapy in childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Moreover, a xenograft mouse model transplanted with primary BCP-ALL cells revealed that neutralized antibody against CD49f improved survival after chemotherapy., Aims: Considering the poor outcomes in Philadelphia chromosome (Ph)-positive ALL treated with conventional chemotherapy without tyrosine kinase inhibitors, we sought to investigate an involvement of the laminin adhesion., Methods and Results: Ph-positive ALL cell lines expressed the highest levels of CD49f among the BCP-ALL cell lines with representative translocations, while CD29 and CD104 were ubiquitously expressed in BCP-ALL cell lines. The association of Ph-positive ALL with high levels of CD49f gene expression was also confirmed in two databases of childhood ALL cohorts. Ph-positive ALL cell lines attached to laminin and their laminin-binding properties were disrupted by blocking antibodies against CD49f and CD29 but not CD104. The cell surface expression of CD49f, but not CD29 and CD104, was downregulated by imatinib treatment in Ph-positive ALL cell lines, but not in their T315I-acquired sublines. Consistently, the laminin-binding properties were disrupted by the imatinib pre-treatment in the Ph-positive ALL cell line, but not in its T315I-acquired subline., Conclusion: BCR::ABL1 plays an essential role in the laminin adhesion of Ph-positive ALL cells through upregulation of CD49f., (© 2024 The Authors. Cancer Reports published by Wiley Periodicals LLC.)
- Published
- 2024
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27. Utility of ASNS gene methylation evaluated with the HPLC method as a pharmacogenomic biomarker to predict asparaginase sensitivity in BCP-ALL.
- Author
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Watanabe A, Miyake K, Yamada Y, Sunamura EI, Yotani T, Kagami K, Kasai S, Tamai M, Harama D, Akahane K, Goi K, Sakaguchi K, Goto H, Kitahara S, and Inukai T
- Subjects
- Humans, Asparaginase genetics, Asparaginase metabolism, Asparaginase therapeutic use, Asparagine genetics, Asparagine metabolism, Asparagine therapeutic use, Chromatography, High Pressure Liquid, Pharmacogenetics, DNA Methylation, Cell Line, Tumor, Aspartate-Ammonia Ligase genetics, Aspartate-Ammonia Ligase metabolism, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics
- Abstract
Asparaginase is an important agent for the treatment of acute lymphoblastic leukaemia (ALL), but it is occasionally associated with severe adverse events. Thus, for safer and more efficacious therapy, a clinical biomarker predicting asparaginase sensitivity is highly anticipated. Asparaginase depletes serum asparagine by deaminating asparagine into aspartic acid, and ALL cells are thought to be sensitive to asparaginase due to reduced asparagine synthetase (ASNS) activity. We have recently shown that allele-specific methylation of the ASNS gene is highly involved in asparaginase sensitivity in B-precursor ALL (BCP-ALL) by using next-generation sequence (NGS) analysis of bisulphite PCR products of the genomic DNA. Here, we sought to confirm the utility of methylation status of the ASNS gene evaluated with high-performance liquid chromatography (HPLC) analysis of bisulphite PCR products for future clinical applications. In the global methylation status of 23 CpG sites at the boundary region of promoter and exon 1 of the ASNS gene, a strong positive correlation was confirmed between the mean percent methylation evaluated with the HPLC method and that with the NGS method in 79 BCP-ALL cell lines (R
2 = 0.85, p = 1.3 × 10-33 ) and in 63 BCP-ALL clinical samples (R2 = 0.84, p = 5.0 × 10-26 ). Moreover, methylation status of the ASNS gene evaluated with the HPLC method was significantly associated with in vitro asparaginase sensitivities as well as gene and protein expression levels of ASNS. These observations indicated that the ASNS gene methylation status evaluated with the HPLC method is a reliable biomarker for predicting the asparaginase sensitivity of BCP-ALL.- Published
- 2023
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28. CRISPR/Cas9-Mediated Induction of Relapse-Specific NT5C2 and PRPS1 Mutations Confers Thiopurine Resistance as a Relapsed Lymphoid Leukemia Model.
- Author
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Nguyen TTT, Tanaka Y, Sanada M, Hosaka M, Tamai M, Kagami K, Komatsu C, Somazu S, Harama D, Kasai S, Watanabe A, Akahane K, Goi K, and Inukai T
- Subjects
- Humans, CRISPR-Cas Systems genetics, Mutation, Recurrence, 5'-Nucleotidase genetics, 5'-Nucleotidase metabolism, 5'-Nucleotidase therapeutic use, Ribose-Phosphate Pyrophosphokinase genetics, Ribose-Phosphate Pyrophosphokinase metabolism, Mercaptopurine pharmacology, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics
- Abstract
6-Mercaptopurine (6-MP) is a key component in maintenance therapy for childhood acute lymphoblastic leukemia (ALL). Recent next-generation sequencing analysis of childhood ALL clarified the emergence of the relapse-specific mutations of the NT5C2 and PRPS1 genes, which are involved in thiopurine metabolism. In this scenario, minor clones of leukemia cells could acquire the 6-MP-resistant phenotype as a result of the NT5C2 or PRPS1 mutation during chemotherapy (including 6-MP treatment) and confer disease relapse after selective expansion. Thus, to establish new therapeutic modalities overcoming 6-MP resistance in relapsed ALL, human leukemia models with NT5C2 and PRPS1 mutations in the intrinsic genes are urgently required. Here, mimicking the initiation process of the above clinical course, we sought to induce two relapse-specific hotspot mutations (R39Q mutation of the NT5C2 gene and S103N mutation of the PRPS1 gene) into a human lymphoid leukemia cell line by homologous recombination (HR) using the CRISPR/Cas9 system. After 6-MP selection of the cells transfected with Cas9 combined with single-guide RNA and donor DNA templates specific for either of those two mutations, we obtained the sublines with the intended NT5C2 -R39Q and PRPS1 -S103N mutation as a result of HR. Moreover, diverse in-frame small insertion/deletions were also confirmed in the 6-MP-resistant sublines at the target sites of the NT5C2 and PRPS1 genes as a result of nonhomologous end joining. These sublines are useful for molecular pharmacological evaluation of the NT5C2 and PRPS1 gene mutations in the 6-MP sensitivity and development of therapy overcoming the thiopurine resistance of leukemia cells. SIGNIFICANCE STATEMENT: Mimicking the initiation process of relapse-specific mutations of the NT5C2 and PRPS1 genes in childhood acute lymphoblastic leukemia treated with 6-mercaptopurine (6-MP), this study sought to introduce NT5C2 -R39Q and PRPS1 -S103N mutations into a human lymphoid leukemia cell line by homologous recombination using the CRISPR/Cas9 system. In the resultant 6-MP-resistant sublines, the intended mutations and diverse in-frame small insertions/deletions were confirmed, indicating that the obtained sublines are useful for molecular pharmacological evaluation of the NT5C2 and PRPS1 gene mutations., (Copyright © 2023 by The American Society for Pharmacology and Experimental Therapeutics.)
- Published
- 2023
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29. Impact of KIR-ligand mismatch on pediatric T-cell acute lymphoblastic leukemia in unrelated cord blood transplantation.
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Kawahara Y, Ishimaru S, Tanaka J, Kako S, Hirayama M, Kanaya M, Ishida H, Sato M, Kobayashi R, Kato M, Goi K, Saito S, Koga Y, Hashii Y, Kato K, Sato A, Atsuta Y, and Sakaguchi H
- Subjects
- Adolescent, Child, Histocompatibility Antigens, Humans, Ligands, Retrospective Studies, T-Lymphocytes, Cord Blood Stem Cell Transplantation, Graft vs Host Disease, Hematopoietic Stem Cell Transplantation, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
- Abstract
Currently, allogeneic hematopoietic stem cell transplantation (allo-HSCT) is considered to be indicated for children and adolescents with high-risk or relapsed T-cell acute lymphoblastic leukemia (T-ALL); however, the outcomes are unsatisfactory. Killer cell immunoglobulin-like receptors (KIRs) are the main receptors on natural killer (NK) cells that play an important role in the graft-versus-leukemia effect after allo-HSCT. In allo-HSCT, when the recipient lacks a donor KIR-ligand (KIR-ligand mismatch in the graft-versus-host [GVH] direction), donor NK cells will be activated against recipient cells. KIR-ligand mismatch in the GVH direction improves outcomes after unrelated cord blood transplantation (UCBT) with acute myeloid leukemia, but the effect in T-ALL is unclear. We evaluated the impact of KIR-ligand mismatch in the GVH direction on the transplantation outcomes of children and adolescents with T-ALL who received UCBT. We conducted a retrospective study using a nationwide registry of the Japanese Society for Transplantation and Cellular Therapy. Patients diagnosed with T-ALL, aged 0 to 19 years, and who underwent first UCBT between 1999 and 2017 were included. A total of 91 patients were included in this study. In all, 23 (25.3%) percent of patients had KIR-ligand mismatch in the GVH direction. The 5-year leukemia-free survival (LFS) and overall survival (OS) rates after UCBT were 65.8% and 69.6%, respectively. In a multivariate analysis, KIR-ligand mismatch in the GVH direction was associated with a significant reduction in the relapse rate (hazard ratio [HR], 0.19; P = .002), resulting in better LFS (HR, 0.18; P =.010) and OS (HR, 0.26; P = .048) without increasing non-relapse mortality (NRM; HR, 1.90; P = .264). The cumulative incidence of GVH disease (GVHD) did not differ between patients with and without KIR-ligand mismatch (grade II-IV acute GVHD, 39.1% versus 36.8%, P = .648, grade III-IV acute GVHD, 13.0% versus 11.8%, P =.857, and chronic GVHD, 26.1% versus 22.9%, P =.736, respectively). Furthermore, acute and chronic GVHD were not associated with good patient outcomes. Notably, no relapse was observed in patients who received KIR-ligand mismatched UCBT in complete remission. KIR-ligand mismatch in the GVH direction improved LFS and decreased relapse rates without increasing NRM in children and adolescents with T-ALL who received UCBT, which was not mediated by GVHD., (Copyright © 2022 The American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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30. Successful treatment of intractable gastrointestinal tract graft-vs-host disease with oral beclomethasone dipropionate in pediatric and young adult patients: Case reports.
- Author
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Akahane K, Watanabe A, Somazu S, Harama D, Shinohara T, Kasai S, Oshiro H, Goi K, Hasuda N, Ozawa C, Sugita K, and Inukai T
- Subjects
- Beclomethasone adverse effects, Beclomethasone therapeutic use, Child, Humans, Young Adult, Gastrointestinal Diseases chemically induced, Gastrointestinal Diseases etiology, Graft vs Host Disease drug therapy, Graft vs Host Disease etiology, Hematopoietic Stem Cell Transplantation adverse effects
- Abstract
Rationale: The gastrointestinal (GI) tract is a common target organ of graft-vs-host disease (GVHD) in hematopoietic stem cell transplantation (HSCT) patients, and GI tract GVHD is often resistant to standard treatments such as corticosteroids. Moreover, longterm use of systemic corticosteroids sometimes induces adverse events such as infection. Beclomethasone dipropionate (BDP) is a potent, topically active corticosteroid, which is metabolized to an active derivative in the intestinal mucosa. Oral BDP therapy is reportedly effective against GI tract GVHD in adult HSCT patients, but its efficacy and safety in pediatric patients remain undefined. Here, we report three pediatric and young adult cases who were treated with oral BDP., Patient Concerns: Three (6-, 7-, and 18-year-old) patients developed stage 2 to 4 lower GI tract GVHD, which was resistant to standard immunosuppressive therapies., Diagnosis: Lower GI tract GVHD in these patients was histopathologically proven by endoscopic biopsy., Interventions: Oral administration of enteric-coated capsules of BDP (3-8 mg/day) was started for the treatment of lower GI tract GVHD., Outcomes: With the introduction of oral BDP therapy, their GI tract symptoms promptly resolved (abdominal pain, within 3-7 days; diarrhea, within 2-3 weeks). Subsequently, systemic immunosuppressive agents such as corticosteroids and mycophenolate mofetil were successfully tapered off. During oral BDP therapy, although cytomegalovirus antigenemia and Acinetobacter Iwoffii sepsis developed in 2 cases, both were curable with conventional treatments. In a young adult case, concomitant BK virus-associated hemorrhagic cystitis resolved after oral BDP was introduced and systemic immunosuppressive agents were reduced. Transient growth restriction was observed in a pediatric case who was treated with oral BDP for approximately 300days., Lessons: Our experiences suggest that oral BDP therapy is an effective approach for GI tract GVHD that is resistant to standard immunosuppressive therapies. Of clinical importance, our case suggests the possibility that oral BDP therapy may improve the immunosuppressive condition in GI tract GVHD patients by contributing to the reduction of systemic immunosuppressive medications as a result of prompt improvement of GI tract GVHD symptoms., Competing Interests: The authors have no conflicts of interests to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)
- Published
- 2022
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31. Association of allele-specific methylation of the ASNS gene with asparaginase sensitivity and prognosis in T-ALL.
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Akahane K, Kimura S, Miyake K, Watanabe A, Kagami K, Yoshimura K, Shinohara T, Harama D, Kasai S, Goi K, Kawai T, Hata K, Kiyokawa N, Koh K, Imamura T, Horibe K, Look AT, Minegishi M, Sugita K, Takita J, and Inukai T
- Subjects
- Alleles, Asparagine genetics, Asparagine metabolism, Cell Line, Tumor, Child, DNA Methylation, Humans, Prognosis, Asparaginase therapeutic use, Aspartate-Ammonia Ligase genetics, Aspartate-Ammonia Ligase metabolism, Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor genetics, Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor metabolism, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma genetics
- Abstract
Asparaginase therapy is a key component of chemotherapy for patients with T-cell acute lymphoblastic leukemia (T-ALL). Asparaginase depletes serum asparagine by deamination into aspartic acid. Normal hematopoietic cells can survive due to asparagine synthetase (ASNS) activity, whereas leukemia cells are supposed to undergo apoptosis due to silencing of the ASNS gene. Because the ASNS gene has a typical CpG island in its promoter, its methylation status in T-ALL cells may be associated with asparaginase sensitivity. Thus, we investigated the significance of ASNS methylation status in asparaginase sensitivity of T-ALL cell lines and prognosis of childhood T-ALL. Sequencing of bisulfite polymerase chain reaction products using next-generation sequencing technology in 22 T-ALL cell lines revealed a stepwise allele-specific methylation of the ASNS gene, in association with an aberrant methylation of a 7q21 imprinted gene cluster. T-ALL cell lines with ASNS hypermethylation status showed significantly higher in vitro l-asparaginase sensitivity in association with insufficient asparaginase-induced upregulation of ASNS gene expression and lower basal ASNS protein expression. A comprehensive analysis of diagnostic samples from pediatric patients with T-ALL in Japanese cohorts (N = 77) revealed that methylation of the ASNS gene was associated with an aberrant methylation of the 7q21 imprinted gene cluster. In pediatric T-ALL patients in Japanese cohorts (n = 75), ASNS hypomethylation status was significantly associated with poor therapeutic outcome, and all cases with poor prognostic SPI1 fusion exclusively exhibited ASNS hypomethylation status. These observations show that ASNS hypomethylation status is associated with asparaginase resistance and is a poor prognostic biomarker in childhood T-ALL., (© 2022 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
- Published
- 2022
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32. NUDT15 polymorphism and NT5C2 and PRPS1 mutations influence thiopurine sensitivity in acute lymphoblastic leukaemia cells.
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Somazu S, Tanaka Y, Tamai M, Watanabe A, Kagami K, Abe M, Harama D, Shinohara T, Akahane K, Goi K, Sugita K, Moriyama T, Yang J, Goto H, Minegishi M, Iwamoto S, Takita J, and Inukai T
- Subjects
- Alleles, Antimetabolites, Antineoplastic pharmacology, Apoptosis genetics, Cell Line, Tumor, Cell Survival genetics, Dose-Response Relationship, Drug, Genotype, Humans, 5'-Nucleotidase genetics, Drug Resistance, Neoplasm genetics, Mercaptopurine pharmacology, Mutation, Polymorphism, Genetic, Pyrophosphatases genetics, Ribose-Phosphate Pyrophosphokinase genetics
- Abstract
In chemotherapy for childhood acute lymphoblastic leukaemia (ALL), maintenance therapy consisting of oral daily mercaptopurine and weekly methotrexate is important. NUDT15 variant genotype is reportedly highly associated with severe myelosuppression during maintenance therapy, particularly in Asian and Hispanic populations. It has also been demonstrated that acquired somatic mutations of the NT5C2 and PRPS1 genes, which are involved in thiopurine metabolism, are detectable in a portion of relapsed childhood ALL. To directly confirm the significance of the NUDT15 variant genotype and NT5C2 and PRPS1 mutations in thiopurine sensitivity of leukaemia cells in the intrinsic genes, we investigated 84 B-cell precursor-ALL (BCP-ALL) cell lines. Three and 14 cell lines had homozygous and heterozygous variant diplotypes of the NUDT15 gene, respectively, while 4 and 2 cell lines that were exclusively established from the samples at relapse had the NT5C2 and PRPS1 mutations, respectively. Both NUDT15 variant genotype and NT5C2 and PRPS1 mutations were significantly associated with DNA-incorporated thioguanine levels after exposure to thioguanine at therapeutic concentration. Considering the continuous exposure during the maintenance therapy, we evaluated in vitro mercaptopurine sensitivity after 7-day exposure. Mercaptopurine concentrations lethal to 50% of the leukaemia cells were comparable to therapeutic serum concentration of mercaptopurine. Both NUDT15 variant genotype and NT5C2 and PRPS1 mutations were significantly associated with mercaptopurine sensitivity in 83 BCP-ALL and 23 T-ALL cell lines. The present study provides direct evidence to support the general principle showing that both inherited genotype and somatically acquired mutation are crucially implicated in the drug sensitivity of leukaemia cells., (© 2021 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)
- Published
- 2021
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33. Epigenetic Modification of Death Receptor Genes for TRAIL and TRAIL Resistance in Childhood B-Cell Precursor Acute Lymphoblastic Leukemia.
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Watanabe A, Miyake K, Akahane K, Goi K, Kagami K, Yagita H, and Inukai T
- Subjects
- Cell Line, Tumor, CpG Islands, Humans, Promoter Regions, Genetic, Receptors, TNF-Related Apoptosis-Inducing Ligand metabolism, TNF-Related Apoptosis-Inducing Ligand metabolism, TNF-Related Apoptosis-Inducing Ligand toxicity, DNA Methylation, Drug Resistance, Neoplasm, Epigenesis, Genetic, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Receptors, TNF-Related Apoptosis-Inducing Ligand genetics
- Abstract
Immunotherapies specific for B-cell precursor acute lymphoblastic leukemia (BCP-ALL), such as anti-CD19 chimeric antigen receptor (CAR) T-cells and blinatumomab, have dramatically improved the therapeutic outcome in refractory cases. In the anti-leukemic activity of those immunotherapies, TNF-related apoptosis-inducing ligand (TRAIL) on cytotoxic T-cells plays an essential role by inducing apoptosis of the target leukemia cells through its death receptors (DR4 and DR5). Since there are CpG islands in the promoter regions, hypermethylation of the DR4 and DR5 genes may be involved in resistance of leukemia cells to immunotherapies due to TRAIL-resistance. We analyzed the DR4 and DR5 methylation status in 32 BCP-ALL cell lines by sequencing their bisulfite PCR products with a next-generation sequencer. The DR4 and DR5 methylation status was significantly associated with the gene and cell-surface expression levels and the TRAIL-sensitivities. In the clinical samples at diagnosis (459 cases in the NOPHO study), both DR4 and DR5 genes were unmethylated in the majority of cases, whereas methylated in several cases with dic(9;20), MLL -rearrangement, and hypodiploidy, suggesting that evaluation of methylation status of the DR4 and DR5 genes might be clinically informative to predict efficacy of immunotherapy in certain cases with such unfavorable karyotypes. These observations provide an epigenetic rational for clinical efficacy of immunotherapy in the vast majority of BCP-ALL cases.
- Published
- 2021
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34. Association of aberrant ASNS imprinting with asparaginase sensitivity and chromosomal abnormality in childhood BCP-ALL.
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Watanabe A, Miyake K, Nordlund J, Syvänen AC, van der Weyden L, Honda H, Yamasaki N, Nagamachi A, Inaba T, Ikawa T, Urayama KY, Kiyokawa N, Ohara A, Kimura S, Kubota Y, Takita J, Goto H, Sakaguchi K, Minegishi M, Iwamoto S, Shinohara T, Kagami K, Abe M, Akahane K, Goi K, Sugita K, and Inukai T
- Subjects
- Animals, Child, Chromosome Aberrations, DNA Methylation genetics, Genomic Imprinting genetics, Humans, Mice, Asparaginase therapeutic use, Aspartate-Ammonia Ligase genetics, Pharmacogenomic Variants genetics, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma genetics
- Abstract
Karyotype is an important prognostic factor in childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL), but the underlying pharmacogenomics remain unknown. Asparaginase is an integral component in current chemotherapy for childhood BCP-ALL. Asparaginase therapy depletes serum asparagine. Normal hematopoietic cells can produce asparagine by asparagine synthetase (ASNS) activity, but ALL cells are unable to synthesize adequate amounts of asparagine. The ASNS gene has a typical CpG island in its promoter. Thus, methylation of the ASNS CpG island could be one of the epigenetic mechanisms for ASNS gene silencing in BCP-ALL. To gain deep insights into the pharmacogenomics of asparaginase therapy, we investigated the association of ASNS methylation status with asparaginase sensitivity. The ASNS CpG island is largely unmethylated in normal hematopoietic cells, but it is allele-specifically methylated in BCP-ALL cells. The ASNS gene is located at 7q21, an evolutionally conserved imprinted gene cluster. ASNS methylation in childhood BCP-ALL is associated with an aberrant methylation of the imprinted gene cluster at 7q21. Aberrant methylation of mouse Asns and a syntenic imprinted gene cluster is also confirmed in leukemic spleen samples from ETV6-RUNX1 knockin mice. In 3 childhood BCP-ALL cohorts, ASNS is highly methylated in BCP-ALL patients with favorable karyotypes but is mostly unmethylated in BCP-ALL patients with poor prognostic karyotypes. Higher ASNS methylation is associated with higher L-asparaginase sensitivity in BCP-ALL through lower ASNS gene and protein expression levels. These observations demonstrate that silencing of the ASNS gene as a result of aberrant imprinting is a pharmacogenetic mechanism for the leukemia-specific activity of asparaginase therapy in BCP-ALL., (© 2020 by The American Society of Hematology.)
- Published
- 2020
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35. Inherited genetic variants associated with glucocorticoid sensitivity in leukaemia cells.
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Shinohara T, Urayama KY, Watanabe A, Akahane K, Goi K, Huang M, Kagami K, Abe M, Sugita K, Okada Y, Goto H, Minegishi M, Iwamoto S, and Inukai T
- Subjects
- Cell Line, Tumor, Dexamethasone pharmacology, Drug Screening Assays, Antitumor, Gene Expression Profiling, Genotype, Humans, Inhibitory Concentration 50, Japan, Pharmacogenetics, Polymorphism, Single Nucleotide, Prednisolone pharmacology, Receptors, Glucocorticoid genetics, Gene Expression Regulation, Leukemic, Genetic Variation, Glucocorticoids pharmacology, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics
- Abstract
Identification of genetic variants associated with glucocorticoids (GC) sensitivity of leukaemia cells may provide insight into potential drug targets and tailored therapy. In the present study, within 72 leukaemic cell lines derived from Japanese patients with B-cell precursor acute lymphoblastic leukaemia (ALL), we conducted genome-wide genotyping of single nucleotide polymorphisms (SNP) and attempted to identify genetic variants associated with GC sensitivity and NR3C1 (GC receptor) gene expression. IC50 measures for prednisolone (Pred) and dexamethasone (Dex) were available using an alamarBlue cell viability assay. IC50 values of Pred showed the strongest association with rs904419 (P = 4.34 × 10
-8 ), located between the FRMD4B and MITF genes. The median IC50 values of prednisolone for cell lines with rs904419 AA (n = 13), AG (n = 31) and GG (n = 28) genotypes were 0.089, 0.139 and 297 µmol/L, respectively. For dexamethasone sensitivity, suggestive association was observed for SNP rs2306888 (P = 1.43 × 10-6 ), a synonymous SNP of the TGFBR3 gene. For NR3C1 gene expression, suggestive association was observed for SNP rs11982167 (P = 6.44 × 10-8 ), located in the PLEKHA8 gene. These genetic variants may affect GC sensitivity of ALL cells and may give rise to opportunities in personalized medicine for effective and safe chemotherapy in ALL patients., (© 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)- Published
- 2020
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36. Association of relapse-linked ARID5B single nucleotide polymorphisms with drug resistance in B-cell precursor acute lymphoblastic leukemia cell lines.
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Tamai M, Huang M, Kagami K, Abe M, Somazu S, Shinohara T, Harama D, Watanabe A, Akahane K, Goi K, Sugita K, Goto H, Minegishi M, Iwamoto S, and Inukai T
- Abstract
Background: The genetic variants of the ARID5B gene have recently been reported to be associated with disease susceptibility and treatment outcome in childhood acute lymphoblastic leukemia (ALL). However, few studies have explored the association of ARID5B with sensitivities to chemotherapeutic agents., Methods: We genotyped susceptibility-linked rs7923074 and rs10821936 as well as relapse-linked rs4948488, rs2893881, and rs6479778 of ARDI5B by direct sequencing of polymerase chain reaction (PCR) products in 72 B-cell precursor-ALL (BCP-ALL) cell lines established from Japanese patients. We also quantified their ARID5B expression levels by real-time reverse transcription PCR, and determined their 50% inhibitory concentration (IC50) values by alamarBlue assays in nine representative chemotherapeutic agents used for ALL treatment., Results: No significant associations were observed in genotypes of the susceptibility-linked single nucleotide polymorphisms (SNPs) and the relapsed-linked SNPs with ARID5B gene expression levels. Of note, IC50 values of vincristine (VCR) (median IC50: 39.6 ng/ml) in 12 cell lines with homozygous genotype of risk allele (C) in the relapse-linked rs4948488 were significantly higher (p = 0.031 in Mann-Whitney U test) than those (1.04 ng/ml) in 60 cell lines with heterozygous or homozygous genotypes of the non-risk allele (T). Furthermore, the IC50 values of mafosfamide [Maf; active metabolite of cyclophosphamide (CY)] and cytarabine (AraC) tended to be associated with the genotype of rs4948488. Similar associations were observed in genotypes of the relapse-linked rs2893881 and rs6479778, but not in those of the susceptibility-linked rs7923074 and rs10821936. In addition, the IC50 values of methotrexate (MTX) were significantly higher (p = 0.023) in 36 cell lines with lower ARID5B gene expression (median IC50: 37.1 ng/ml) than those in the other 36 cell lines with higher expression (16.9 ng/ml)., Conclusion: These observations in 72 BCP-ALL cell lines suggested that the risk allele of the relapse-linked SNPs of ARID5B may be involved in a higher relapse rate because of resistance to chemotherapeutic agents such as VCR, CY, and AraC. In addition, lower ARID5B gene expression may be associated with MTX resistance.
- Published
- 2020
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37. Resistance of t(17;19)-acute lymphoblastic leukemia cell lines to multiagents in induction therapy.
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Watanabe A, Inukai T, Kagami K, Abe M, Takagi M, Fukushima T, Fukushima H, Nanmoku T, Terui K, Ito T, Toki T, Ito E, Fujimura J, Goto H, Endo M, Look T, Kamps M, Minegishi M, Takita J, Inaba T, Takahashi H, Ohara A, Harama D, Shinohara T, Somazu S, Oshiro H, Akahane K, Goi K, and Sugita K
- Subjects
- ATP Binding Cassette Transporter, Subfamily B, Member 1 genetics, ATP Binding Cassette Transporter, Subfamily B, Member 1 metabolism, Alleles, Animals, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Cell Line, Disease Models, Animal, Dose-Response Relationship, Drug, Female, Gene Frequency, Genotype, Humans, Immunophenotyping, Mice, Neoplasm, Residual diagnosis, Neoplasm, Residual genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma diagnosis, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Chromosomes, Human, Pair 17, Chromosomes, Human, Pair 19, Drug Resistance, Neoplasm genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Translocation, Genetic
- Abstract
t(17;19)(q21-q22;p13), responsible for TCF3-HLF fusion, is a rare translocation in childhood B-cell precursor acute lymphoblastic leukemia(BCP-ALL). t(1;19)(q23;p13), producing TCF3-PBX1 fusion, is a common translocation in childhood BCP-ALL. Prognosis of t(17;19)-ALL is extremely poor, while that of t(1;19)-ALL has recently improved dramatically in intensified chemotherapy. In this study, TCF3-HLF mRNA was detectable at a high level during induction therapy in a newly diagnosed t(17;19)-ALL case, while TCF3-PBX1 mRNA was undetectable at the end of induction therapy in most newly diagnosed t(1;19)-ALL cases. Using 4 t(17;19)-ALL and 16 t(1;19)-ALL cell lines, drug response profiling was analyzed. t(17;19)-ALL cell lines were found to be significantly more resistant to vincristine (VCR), daunorubicin (DNR), and prednisolone (Pred) than t(1;19)-ALL cell lines. Sensitivities to three (Pred, VCR, and l-asparaginase [l-Asp]), four (Pred, VCR, l-Asp, and DNR) and five (Pred, VCR, l-Asp, DNR, and cyclophosphamide) agents, widely used in induction therapy, were significantly poorer for t(17;19)-ALL cell lines than for t(1;19)-ALL cell lines. Consistent with poor responses to VCR and DNR, gene and protein expression levels of P-glycoprotein (P-gp) were higher in t(17;19)-ALL cell lines than in t(1;19)-ALL cell lines. Inhibitors for P-gp sensitized P-gp-positive t(17;19)-ALL cell lines to VCR and DNR. Knockout of P-gp by CRISPRCas9 overcame resistance to VCR and DNR in the P-gp-positive t(17;19)-ALL cell line. A combination of cyclosporine A with DNR prolonged survival of NSG mice inoculated with P-gp-positive t(17;19)-ALL cell line. These findings indicate involvement of P-gp in resistance to VCR and DNR in Pgp positive t(17;19)-ALL cell lines. In all four t(17;19)-ALL cell lines, RAS pathway mutation was detected. Furthermore, among 16 t(1;19)-ALL cell lines, multiagent resistance was usually observed in the cell lines with RAS pathway mutation in comparison to those without it, suggesting at least a partial involvement of RAS pathway mutation in multiagent resistance of t(17;19)-ALL., (© 2019 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)
- Published
- 2019
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38. Autologous Stem Cell Rescue for Graft Failure of Second Allogeneic Stem Cell Transplant After Engraftment of Primary Allogeneic Transplant.
- Author
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Watanabe A, Inukai T, Akahane K, Somazu S, Oshiro H, Goi K, Koizumi K, Harii N, Matsuda K, and Sugita K
- Subjects
- Bone Neoplasms pathology, Child, Fatal Outcome, Hematopoiesis, Humans, Male, Osteosarcoma secondary, Reoperation, Transplantation Conditioning, Transplantation, Autologous, Transplantation, Homologous, Treatment Failure, Bone Neoplasms surgery, Hematopoietic Stem Cell Transplantation adverse effects, Osteosarcoma surgery, Peripheral Blood Stem Cell Transplantation, Tibia pathology
- Abstract
Here, we describe a case of primary graft failure with severe sepsis in a boy who experienced frequent relapses of osteosarcoma. The patient had undergone haploidentical bone marrow transplant after engraftment of unrelated cord blood transplant performed 10 months earlier. Considering his severe condition, we transfused autologous peripheral stem cells along with a single dose of etoposide (50 mg/m2). Granulocyte engraftment was confirmed on human leukocyte antigen-microsatellite analysis of bone marrow on day 14. Although the patient died due to respiratory failure, transfusion of autologous hematopoietic stem cells is a reasonable rescue option for graft failure even in patients whose background hematopoiesis is reconstituted by a first donor.
- Published
- 2019
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39. De Novo Mutations Activating Germline TP53 in an Inherited Bone-Marrow-Failure Syndrome.
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Toki T, Yoshida K, Wang R, Nakamura S, Maekawa T, Goi K, Katoh MC, Mizuno S, Sugiyama F, Kanezaki R, Uechi T, Nakajima Y, Sato Y, Okuno Y, Sato-Otsubo A, Shiozawa Y, Kataoka K, Shiraishi Y, Sanada M, Chiba K, Tanaka H, Terui K, Sato T, Kamio T, Sakaguchi H, Ohga S, Kuramitsu M, Hamaguchi I, Ohara A, Kanno H, Miyano S, Kojima S, Ishiguro A, Sugita K, Kenmochi N, Takahashi S, Eto K, Ogawa S, and Ito E
- Subjects
- Adolescent, Adult, Agammaglobulinemia genetics, Anemia, Diamond-Blackfan genetics, Animals, Child, Preschool, Erythrocytes pathology, Female, Growth Disorders genetics, Humans, Induced Pluripotent Stem Cells pathology, Infant, Infant, Newborn, Male, Mice, Middle Aged, Young Adult, Zebrafish, Bone Marrow pathology, Bone Marrow Diseases genetics, Germ Cells pathology, Mutation genetics, Tumor Suppressor Protein p53 genetics
- Abstract
Inherited bone-marrow-failure syndromes (IBMFSs) include heterogeneous genetic disorders characterized by bone-marrow failure, congenital anomalies, and an increased risk of malignancy. Many lines of evidence have suggested that p53 activation might be central to the pathogenesis of IBMFSs, including Diamond-Blackfan anemia (DBA) and dyskeratosis congenita (DC). However, the exact role of p53 activation in each clinical feature remains unknown. Here, we report unique de novo TP53 germline variants found in two individuals with an IBMFS accompanied by hypogammaglobulinemia, growth retardation, and microcephaly mimicking DBA and DC. TP53 is a tumor-suppressor gene most frequently mutated in human cancers, and occasional germline variants occur in Li-Fraumeni cancer-predisposition syndrome. Most of these mutations affect the core DNA-binding domain, leading to compromised transcriptional activities. In contrast, the variants found in the two individuals studied here caused the same truncation of the protein, resulting in the loss of 32 residues from the C-terminal domain (CTD). Unexpectedly, the p53 mutant had augmented transcriptional activities, an observation not previously described in humans. When we expressed this mutant in zebrafish and human-induced pluripotent stem cells, we observed impaired erythrocyte production. These findings together with close similarities to published knock-in mouse models of TP53 lacking the CTD demonstrate that the CTD-truncation mutations of TP53 cause IBMFS, providing important insights into the previously postulated connection between p53 and IBMFSs., (Copyright © 2018 American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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40. T315I mutation of BCR-ABL1 into human Philadelphia chromosome-positive leukemia cell lines by homologous recombination using the CRISPR/Cas9 system.
- Author
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Tamai M, Inukai T, Kojika S, Abe M, Kagami K, Harama D, Shinohara T, Watanabe A, Oshiro H, Akahane K, Goi K, Sugihara E, Nakada S, and Sugita K
- Subjects
- Cell Line, Tumor, Drug Resistance, Neoplasm genetics, Humans, Imatinib Mesylate pharmacology, K562 Cells, Leukemia, Myelogenous, Chronic, BCR-ABL Positive drug therapy, Leukemia, Myelogenous, Chronic, BCR-ABL Positive pathology, Phthalazines pharmacology, Piperazines pharmacology, Poly(ADP-ribose) Polymerase Inhibitors pharmacology, Protein Kinase Inhibitors pharmacology, Pyrimidines pharmacology, Schiff Bases pharmacology, CRISPR-Cas Systems, Fusion Proteins, bcr-abl genetics, Homologous Recombination, Leukemia, Myelogenous, Chronic, BCR-ABL Positive genetics, Mutation
- Abstract
In many cancers, somatic mutations confer tumorigenesis and drug-resistance. The recently established clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system is a potentially elegant approach to functionally evaluate mutations in cancers. To reproduce mutations by homologous recombination (HR), the HR pathway must be functional, but DNA damage repair is frequently impaired in cancers. Imatinib is a tyrosine kinase inhibitor for BCR-ABL1 in Philadelphia chromosome-positive (Ph+) leukemia, and development of resistance due to kinase domain mutation is an important issue. We attempted to introduce the T315I gatekeeper mutation into three Ph+ myeloid leukemia cell lines with a seemingly functional HR pathway due to resistance to the inhibitor for poly (ADP) ribose polymerase1. Imatinib-resistant sublines were efficiently developed by the CRISPR/Cas9 system after short-term selection with imatinib; resulting sublines acquired the T315I mutation after HR. Thus, the usefulness of CRISPR/Cas9 system for functional analysis of somatic mutations in cancers was demonstrated.
- Published
- 2018
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41. Clofarabine exerts antileukemic activity against cytarabine-resistant B-cell precursor acute lymphoblastic leukemia with low deoxycytidine kinase expression.
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Huang M, Inukai T, Miyake K, Tanaka Y, Kagami K, Abe M, Goto H, Minegishi M, Iwamoto S, Sugihara E, Watanabe A, Somazu S, Shinohara T, Oshiro H, Akahane K, Goi K, and Sugita K
- Subjects
- Amino Acid Sequence, Apoptosis drug effects, Apoptosis genetics, Base Sequence, CRISPR-Cas Systems, Cell Line, Tumor, Cell Survival drug effects, Cell Survival genetics, DNA Methylation, Dose-Response Relationship, Drug, Exons, Gene Expression Regulation, Neoplastic, Gene Knockout Techniques, Humans, Mutation, Polymorphism, Genetic, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma pathology, Promoter Regions, Genetic, Antimetabolites, Antineoplastic pharmacology, Clofarabine pharmacology, Deoxycytidine Kinase genetics, Drug Resistance, Neoplasm drug effects, Drug Resistance, Neoplasm genetics, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma genetics
- Abstract
Cytosine arabinoside (Ara-C) is one of the key drugs for the treatment of acute myeloid leukemia. It is also used for consolidation therapy of acute lymphoblastic leukemia (ALL). Ara-C is a deoxyadenosine analog and is phosphorylated to form cytosine arabinoside triphosphate (Ara-CTP) as an active form. In the first step of the metabolic pathway, Ara-C is phosphorylated to Ara-CMP by deoxycytidine kinase (DCK). However, the current cumulative evidence in the association of the Ara-C sensitivity in ALL appears inconclusive. We analyzed various cell lines for the possible involvement of DCK in the sensitivities of B-cell precursor ALL (BCP-ALL) to Ara-C. Higher DCK expression was associated with higher Ara-C sensitivity. DCK knockout by genome editing with a CRISPR-Cas9 system in an Ara-C-sensitive-ALL cell line induced marked resistance to Ara-C, but not to vincristine and daunorubicin, indicating the involvement of DCK expression in the Ara-C sensitivity of BCP-ALL. DCK gene silencing due to the hypermethylation of a CpG island and reduced DCK activity due to a nonsynonymous variant allele were not associated with Ara-C sensitivity. Clofarabine is a second-generation deoxyadenosine analog rationally synthesized to improve stability and reduce toxicity. The IC50 of clofarabine in 79 BCP-ALL cell lines was approximately 20 times lower than that of Ara-C. In contrast to Ara-C, although the knockout of DCK induced marked resistance to clofarabine, sensitivity to clofarabine was only marginally associated with DCK gene expression level, suggesting a possible efficacy of clofarabine for BCP-ALL that shows relative Ara-C resistance due to low DCK expression., (© 2018 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)
- Published
- 2018
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42. Anti-leukemic activity of bortezomib and carfilzomib on B-cell precursor ALL cell lines.
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Takahashi K, Inukai T, Imamura T, Yano M, Tomoyasu C, Lucas DM, Nemoto A, Sato H, Huang M, Abe M, Kagami K, Shinohara T, Watanabe A, Somazu S, Oshiro H, Akahane K, Goi K, Kikuchi J, Furukawa Y, Goto H, Minegishi M, Iwamoto S, and Sugita K
- Subjects
- B-Lymphocytes metabolism, Cell Line, Tumor, Humans, Antineoplastic Agents pharmacology, B-Lymphocytes drug effects, Bortezomib therapeutic use, Oligopeptides therapeutic use
- Abstract
Prognosis of childhood acute lymphoblastic leukemia (ALL) has been dramatically improved. However, prognosis of the cases refractory to primary therapy is still poor. Recent phase 2 study on the efficacy of combination chemotherapy with bortezomib (BTZ), a proteasome inhibitor, for refractory childhood ALL demonstrated favorable clinical outcomes. However, septic death was observed in over 10% of patients, indicating the necessity of biomarkers that could predict BTZ sensitivity. We investigated in vitro BTZ sensitivity in a large panel of ALL cell lines that acted as a model system for refractory ALL, and found that Philadelphia chromosome-positive (Ph+) ALL, IKZF1 deletion, and biallelic loss of CDKN2A were associated with favorable response. Even in Ph-negative ALL cell lines, IKZF1 deletion and bilallelic loss of CDKN2A were independently associated with higher BTZ sensitivity. BTZ showed only marginal cross-resistance to four representative chemotherapeutic agents (vincristine, dexamethasone, l-asparaginase, and daunorubicin) in B-cell precursor-ALL cell lines. To improve the efficacy and safety of proteasome inhibitor combination chemotherapy, we also analyzed the anti-leukemic activity of carfilzomib (CFZ), a second-generation proteasome inhibitor, as a substitute for BTZ. CFZ showed significantly higher activity than BTZ in the majority of ALL cell lines except for the P-glycoprotein-positive t(17;19) ALL cell lines, and IKZF1 deletion was also associated with a favorable response to CFZ treatment. P-glycoprotein inhibitors effectively restored the sensitivity to CFZ, but not BTZ, in P-glycoprotein-positive t(17;19) ALL cell lines. P-glycoprotein overexpressing ALL cell line showed a CFZ-specific resistance, while knockout of P-glycoprotein by genome editing with a CRISPR/Cas9 system sensitized P-glycoprotein-positive t(17;19) ALL cell line to CFZ. These observations suggested that IKZF1 deletion could be a useful biomarker to predict good sensitivity to CFZ and BTZ, and that CFZ combination chemotherapy may be a new therapeutic option with higher anti-leukemic activity for refractory ALL that contain P-glycoprotein-negative leukemia cells.
- Published
- 2017
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43. Inflammatory mediator ultra-low-molecular-weight hyaluronan triggers necrosis of B-precursor leukemia cells with high surface CD44 expression.
- Author
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Kasai S, Furuichi Y, Ando N, Kagami K, Abe M, Nakane T, Goi K, Inukai T, Saitoh S, Ohno S, Okazaki S, Nagano O, Saya H, and Sugita K
- Subjects
- Biological Transport drug effects, Cell Line, Tumor, Cell Survival drug effects, Child, Child, Preschool, Female, Gene Expression, HMGB1 Protein metabolism, Humans, Hyaluronan Receptors metabolism, Hyaluronic Acid antagonists & inhibitors, Imidazoles pharmacology, Indoles pharmacology, Infant, Male, Molecular Weight, Necrosis chemically induced, Necrosis metabolism, Necrosis pathology, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma metabolism, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma pathology, Precursor Cell Lymphoblastic Leukemia-Lymphoma metabolism, Precursor Cell Lymphoblastic Leukemia-Lymphoma pathology, Precursor Cells, B-Lymphoid drug effects, Precursor Cells, B-Lymphoid metabolism, Precursor Cells, B-Lymphoid pathology, Reactive Oxygen Species agonists, Reactive Oxygen Species metabolism, Thymidine metabolism, Hyaluronan Receptors genetics, Hyaluronic Acid pharmacology, Necrosis genetics, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics
- Abstract
Acute lymphoblastic leukemia (ALL) with mixed lineage leukemia (MLL) gene rearrangements (MLL+ALL) has a dismal prognosis and is characterized by high surface CD44 expression. Known that CD44 has the specific binding sites for a natural ligand hyaluronan (HA), we investigated biological effects of HA with different molecular sizes on MLL+ALL cell lines, and found that the addition of ultra-low-molecular-weight (ULMW)-HA strongly suppressed their thymidine uptakes. The MLL+ALL cell line lacking surface CD44 expression established by genome editing showed no suppression of thymidine uptake. Surface CD44-high B-precursor ALL cell lines other than MLL+, but not T-ALL cell lines, were also suppressed in their thymidine uptakes. The inhibition of thymidine uptakes was because of induction of cell death, but dead cells lacked features of apoptosis on cytospin smears and flow cytometric analysis. The cell death was neither blocked by pan-caspase inhibitor nor autophagy inhibitor, but was completely blocked by necrosis inhibitor necrostatin-1. Necrotic cell death was further supported by a marked release of a high-mobility protein group B1 and morphological changes on transmission electron microscopy. Elevation of intracellular reactive oxygen species production suggested a role for inducing this necrotic cell death. ULMW-HA-triggered cell death was similarly demonstrated in surface CD44-high primary B-precursor leukemia cells. Assuming that ULMW-HA is abundantly secreted at the site of infection and inflammation, this study sheds light on understanding the mechanism of a transient inflammation-associated remission of leukemia. Further, the CD44-targeting may become an effective approach in future for the treatment of refractory B-precursor ALL by its capability of predominantly eradicating CD44-high leukemia-initiating cells.
- Published
- 2017
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44. Specific Antileukemic Activity of PD0332991, a CDK4/6 Inhibitor, against Philadelphia Chromosome-Positive Lymphoid Leukemia.
- Author
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Nemoto A, Saida S, Kato I, Kikuchi J, Furukawa Y, Maeda Y, Akahane K, Honna-Oshiro H, Goi K, Kagami K, Kimura S, Sato Y, Okabe S, Niwa A, Watanabe K, Nakahata T, Heike T, Sugita K, and Inukai T
- Subjects
- Animals, Apoptosis drug effects, Cell Cycle Checkpoints drug effects, Cell Line, Tumor, Cell Proliferation drug effects, Cyclin D2 genetics, Cyclin D2 metabolism, Cyclin-Dependent Kinase Inhibitor p16 metabolism, Disease Models, Animal, Dose-Response Relationship, Drug, Fusion Proteins, bcr-abl genetics, Humans, Mice, Mutation, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma metabolism, Precursor Cell Lymphoblastic Leukemia-Lymphoma mortality, Precursor Cell Lymphoblastic Leukemia-Lymphoma pathology, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Cyclin-Dependent Kinase 4 antagonists & inhibitors, Cyclin-Dependent Kinase 6 antagonists & inhibitors, Piperazines pharmacology, Protein Kinase Inhibitors pharmacology, Pyridines pharmacology
- Abstract
S-phase progression of the cell cycle is accelerated in tumors through various genetic abnormalities, and, thus, pharmacologic inhibition of altered cell-cycle progression would be an effective strategy to control tumors. In the current study, we analyzed the antileukemic activity of three available small molecules targeting CDK4/CDK6 against lymphoid crisis of chronic myeloid leukemia (CML-LC) and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL), and found that all three molecules showed specific activities against leukemic cell lines derived from CML-LC and Ph(+) ALL. In particular, PD0332991 exhibited extremely high antileukemic activity against CML-LC and Ph(+) ALL cell lines in the nanomolar range by the induction of G0-G1 arrest and partially cell death through dephosphorylation of pRb and downregulation of the genes that are involved in S-phase transition. As an underlying mechanism for favorable sensitivity to the small molecules targeting CDK4/CDK6, cell-cycle progression of Ph(+) lymphoid leukemia cells was regulated by transcriptional and posttranscriptional modulation of CDK4 as well as Cyclin D2 gene expression under the control of BCR-ABL probably through the PI3K pathway. Consistently, the gene expression level of Cyclin D2 in Ph(+) lymphoid leukemia cells was significantly higher than that in Ph(-) lymphoid leukemia cells. Of note, three Ph(+) ALL cell lines having the T315I mutation also showed sensitivity to PD0332991. In a xenograft model, PD0332991, but not imatinib, suppressed dissemination of Ph(+) ALL having the T315I mutation and prolonged survival, demonstrating that this reagent would be a new therapeutic modality for relapsed CML-LC and Ph(+) ALL patients after treatment with tyrosine kinase inhibitors., (©2015 American Association for Cancer Research.)
- Published
- 2016
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- View/download PDF
45. Low-loss high-speed silicon IQ modulator for QPSK/DQPSK in C and L bands.
- Author
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Goi K, Oka A, Kusaka H, Terada Y, Ogawa K, Liow TY, Tu X, Lo GQ, and Kwong DL
- Subjects
- Equipment Design, Nonlinear Dynamics, Signal-To-Noise Ratio, Fiber Optic Technology instrumentation, Signal Processing, Computer-Assisted instrumentation, Silicon, Telecommunications instrumentation
- Abstract
A low-loss high-speed silicon in-phase (I) quadrature (Q) modulator is designed, fabricated and characterized. The fabricated IQ modulator has a low passive optical loss of 9 dB in C and L bands. Using the modulator, differential quadrature phase-shift keying (DQPSK) transmission at 44.6 Gb/s with differential detection is confirmed with an optical signal-to-noise ratio (OSNR) of 16.3 dB for a bit error rate (BER) of 10(-3) and a dispersion tolerance of -96 to 107 ps/nm. Moreover, in digital coherent detection, quadrature phase-shift keying (QPSK) up to 64 Gb/s are achieved with an OSNR of 11.6-11.8 dB for a BER of 10(-2) at 1530, 1550, and 1610 nm.
- Published
- 2014
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46. 11-Gb/s 80-km transmission performance of zero-chirp silicon Mach-Zehnder modulator.
- Author
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Goi K, Oda K, Kusaka H, Terada Y, Ogawa K, Liow TY, Tu X, Lo GQ, and Kwong DL
- Abstract
The 11-Gbps 80-km transmission performance of a zero-chirp silicon Mach-Zehnder modulator has been characterized. The zero-chirp characteristic of the silicon modulator is confirmed in the constellation measurement, and gives high tolerance both for positive and negative chromatic dispersion. A low-dispersion-penalty transmission up to 80 km using the 11-Gbps non return-to-zero on-off-keying format is confirmed via bit-error-rate measurements with a performance comparable to that of a commercial lithium-niobate modulator. The dispersion tolerance at 2-dB power penalty for a bit-error-rate of 10(-3) is more than ± 950 ps/nm. Further, 22.3-Gbps binary phase-shift-keying is demonstrated, and the back-to-back power penalty with respect to the lithium-niobate modulator is less than 0.5 dB.
- Published
- 2012
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47. Extensive gene deletions in Japanese patients with Diamond-Blackfan anemia.
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Kuramitsu M, Sato-Otsubo A, Morio T, Takagi M, Toki T, Terui K, Wang R, Kanno H, Ohga S, Ohara A, Kojima S, Kitoh T, Goi K, Kudo K, Matsubayashi T, Mizue N, Ozeki M, Masumi A, Momose H, Takizawa K, Mizukami T, Yamaguchi K, Ogawa S, Ito E, and Hamaguchi I
- Subjects
- Anemia, Diamond-Blackfan ethnology, Anemia, Diamond-Blackfan pathology, Asian People genetics, Child, Preschool, DNA Mutational Analysis methods, Female, Genetic Association Studies, Humans, Infant, Infant, Newborn, Japan, Male, Microarray Analysis methods, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, Reproducibility of Results, Anemia, Diamond-Blackfan genetics, Gene Deletion, Ribosomal Proteins genetics
- Abstract
Fifty percent of Diamond-Blackfan anemia (DBA) patients possess mutations in genes coding for ribosomal proteins (RPs). To identify new mutations, we investigated large deletions in the RP genes RPL5, RPL11, RPL35A, RPS7, RPS10, RPS17, RPS19, RPS24, and RPS26. We developed an easy method based on quantitative-PCR in which the threshold cycle correlates to gene copy number. Using this approach, we were able to diagnose 7 of 27 Japanese patients (25.9%) possessing mutations that were not detected by sequencing. Among these large deletions, similar results were obtained with 6 of 7 patients screened with a single nucleotide polymorphism array. We found an extensive intragenic deletion in RPS19, including exons 1-3. We also found 1 proband with an RPL5 deletion, 1 patient with an RPL35A deletion, 3 with RPS17 deletions, and 1 with an RPS19 deletion. In particular, the large deletions in the RPL5 and RPS17 alleles are novel. All patients with a large deletion had a growth retardation phenotype. Our data suggest that large deletions in RP genes comprise a sizable fraction of DBA patients in Japan. In addition, our novel approach may become a useful tool for screening gene copy numbers of known DBA genes.
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- 2012
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48. Silicon Mach-Zehnder modulator of extinction ratio beyond 10 dB at 10.0-12.5 Gbps.
- Author
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Ogawa K, Goi K, Tan YT, Liow TY, Tu X, Fang Q, Lo GQ, and Kwong DL
- Abstract
Silicon Mach-Zehnder modulators with reduced series resistance in lateral PN junction rib-waveguide phase shifters for enhanced high-speed response are fabricated and characterized. Extinction ratio higher than 10 dB is obtained at 10.3-11.7 Gbps with mask margins of 27% (10.3-Gbps 10GBE), 16% (10.7-Gbps STM-64/OC-192) and 10% (11.3-Gbps STM-64/OC-192) in eye-diagram measurements incorporating mask tests using a RF cut-off filter. In unfiltered eye-diagram measurements without mask tests, extinction ratio higher than 13 dB is obtained at 10.0-12.5 Gbps. The silicon modulators reveal high-speed performance comparable with that of lithium-niobate modulators in high-speed optical fiber telecommunications., (© 2011 Optical Society of America)
- Published
- 2011
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49. Resistance of T-cell acute lymphoblastic leukemia to tumor necrosis factor--related apoptosis-inducing ligand-mediated apoptosis.
- Author
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Akahane K, Inukai T, Zhang X, Hirose K, Kuroda I, Goi K, Honna H, Kagami K, Nakazawa S, Endo K, Kubota T, Yagita H, Koyama-Okazaki T, and Sugita K
- Subjects
- Cell Line, Tumor, Cells, Cultured, DNA Methylation drug effects, Dose-Response Relationship, Drug, Drug Resistance, Flow Cytometry, Gene Expression drug effects, Humans, Immunoblotting, Jurkat Cells, Luciferases genetics, Luciferases metabolism, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma metabolism, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma pathology, Promoter Regions, Genetic genetics, Receptors, TNF-Related Apoptosis-Inducing Ligand genetics, Reverse Transcriptase Polymerase Chain Reaction, fas Receptor metabolism, Apoptosis drug effects, Cell Proliferation drug effects, Fas Ligand Protein pharmacology, TNF-Related Apoptosis-Inducing Ligand pharmacology
- Abstract
Objective: Cytotoxic ligands are involved in tumor immunity and graft-vs.-leukemia effect after allogeneic stem cell transplantation for leukemia. To clarify the susceptibility of T-cell acute lymphoblastic leukemia (T-ALL) to tumor immunity, sensitivity to recombinant human soluble Fas ligand (rhsFasL) and tumor necrosis factor-related apoptosis-inducing ligand (rhsTRAIL) was determined., Materials and Methods: Sensitivity to rhsFasL and rhsTRAIL and cell surface expression of their receptors were tested in T-ALL cell lines (n = 7) and patients' samples (n = 17) and compared with those in B-precursor ALL cell lines (n = 30). Expression of components of the death-inducing signaling complex and the TRAIL receptor genes (DR4/DR5), and the methylation status and promoter activity of the DR4/DR5 gene were tested in T-ALL cell lines., Results: T-ALL cell lines showed higher level of Fas expression and higher sensitivity to rhsFasL than did B-precursor ALL cell lines. Despite comparable expression of components of death-inducing signaling complex, cell lines and patients' samples of T-ALL showed TRAIL-resistance associated with low cell surface expression of DR4/DR5. Gene expression of DR4/DR5 in T-ALL cell lines was significantly lower than that in B-precursor ALL cell lines, and the methylation status of the gene promoter in T-ALL cell lines was associated with the gene expression level at least for DR4. The demethylating agent, 5-aza 2'deoxycytidine, upregulated the gene expression of DR4/DR5, but was insufficient for their surface expression due to low basal promoter activity., Conclusions: In contrast to higher sensitivity to FasL, T-ALL showed resistance to TRAIL, which might be responsible for resistance to TRAIL-mediated cellular immunity., (Copyright © 2010 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc. All rights reserved.)
- Published
- 2010
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50. Fertility recovery and pregnancy after allogeneic hematopoietic stem cell transplantation in Fanconi anemia patients.
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Nabhan SK, Bitencourt MA, Duval M, Abecasis M, Dufour C, Boudjedir K, Rocha V, Socié G, Passweg J, Goi K, Sanders J, Snowden J, Yabe H, Pasquini R, and Gluckman E
- Subjects
- Fanconi Anemia therapy, Female, Humans, Infant, Newborn, Male, Pregnancy, Retrospective Studies, Transplantation, Homologous, Fanconi Anemia complications, Hematopoietic Stem Cell Transplantation methods, Infertility etiology, Recovery of Function
- Abstract
Reduced fertility is one clinical manifestation among other well known Fanconi anemia features. Most recipients of allogeneic hematopoietic stem cell transplantation suffer from secondary infertility owing to gonadal damage from myeloablative conditioning. In order to evaluate the rate of pregnancy in Fanconi anemia transplanted patients, we performed a retrospective analysis of female patients transplanted in 15 centers from 1976 to 2008. Among 578 transplanted Fanconi anemia patients, we identified 285 transplanted females of whom 101 patients were aged 16 years or over. Ten became pregnant (4 twice). Before hematopoietic stem cell transplantation all had confirmed Fanconi anemia diagnosis. Median age at transplantation was 12 years (range 5-17 years). Conditioning regimen consisted of cyclophosphamide with or without irradiation. During follow up, 5 of 10 patients presented signs of ovarian failure. Among those, 2 patients spontaneously recovered regular menses, and 3 received hormonal replacement therapy. Pregnancy occurred from four to 17 years after hematopoietic stem cell transplantation. Three patients had preterm deliveries, one patient had a hysterectomy for bleeding. All 14 newborns had normal growth and development without congenital diseases. In conclusion, recovery of normal ovarian function and a viable pregnancy is a realistic but relatively rare possibility even in Fanconi anemia patients following hematopoietic stem cell transplantation. Mechanisms of fertility recovery are discussed.
- Published
- 2010
- Full Text
- View/download PDF
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