Objective： To analyze Fms-like tyrosine kinase 3 （FLT3）/internal-tandem duplications （ITD） mutations in various kinds of hematologic malignancy patients. Methods： FLT3/ITD gene mutations were detected by polymer...Objective： To analyze Fms-like tyrosine kinase 3 （FLT3）/internal-tandem duplications （ITD） mutations in various kinds of hematologic malignancy patients. Methods： FLT3/ITD gene mutations were detected by polymerase chain reaction （PCR） in 103 acute myeloid leukemia （AML） cases, 63 acute lymphocytic leukemia （ALL） cases, 53 chronic myelogenous leukemia （CML） cases in chronic phase （CML-CP）, 34 CML cases in blast crisis （CML-BC）, 11 chronic lymphatic leukemia （CLL） cases, 36 myelodysplastic syndrome （MDS） cases, 9 multiple myeloma （MM） cases and 13 non-hodgkin＇s lymphoma （NHL） cases with marrow infiltration. Results： The expressions of FLT3/ITD gene mutations were detected in 22.3% AML cases, in 6.5% CML-BC cases, in 5.6% MDS cases and in 2.6% ALL cases. The two ALL cases with FLT3/ITD mutation were diagnosed as ALL-L2 with morphology and both with myeloid antigen expression, but finally were diagnosed as acute mixed-lineage leukemia after immunology examination. FLT3/ITD gene mutations were not detected in CML-CP, MM, NHL and CLL cases. In the 23 AML patients with FLT3/ITD gene mutation, including 2 of 8 M1 （2.5%）, 8 of 33 M2 （24.2%）, 7 of 24 M3 （29.3%）, 2 of 11 M4 （18.2%）, 3 of 21 M5 （14.3%）, 1 of 5 M6 （20%）, and 0 of 1 M7 cases, and there were no significant differences in the positive rates of FLT3/ITD mutations between the FAB subtypes （P 〉 0.05）. Statistical analyses showed that in AML patients, FLT3/ITD was associated with a higher peripheral blood white cell （WBC） counts [（41.23 ± 32.56） x 109/L vs （11.36 ± 9.89） × 10^9/L （P 〈 0.01 ）], higher percentage of bone marrow blast cells [（72.78 ± 21.79）% vs （51.26 ± 20.78）% （P 〈 0.05）], and higher cumulative relapse rates （63.6% vs 27.7%, P 〈 0.025） than those negative. Conclusion： FLT3/ITD gene mutation mainly occurred in AML patients, and might be a strong prognostic factor which was associated with high peripheral WBC counts, bone marrow blast cell proportion and a increased relapse risk in AML. Detection of FLT3/ITD gene mutation might provide insights to explore a more accurate genotyping of leukemia, differential diagnosis between AML and ALL, subdivide risk level in AML and estimate prognosis of leukemia.展开更多
FMS-like tyrosine kinase 3(FLT3)mutation is strongly associated with poor prognosis in acute myeloid leukemia(AML).Though many FLT3 inhibitors have been developed for clinical application with 34%-56%complete remissio...FMS-like tyrosine kinase 3(FLT3)mutation is strongly associated with poor prognosis in acute myeloid leukemia(AML).Though many FLT3 inhibitors have been developed for clinical application with 34%-56%complete remission rate,patients would develop resistance sooner or later after initial response to tyrosine kinase inhibitors(TKIs),such as gilteritinib.And increasing studies have shown that several resistance related mutations of FLT3 emerged during the AML progression.Thus,further investigation is warranted for these FLT3mu,AML patients to achieve a better treatment outcome.4-Hydroxyphenyl retinamide(4-HPR)has been investigated extensively in animal models and clinical trials as an anticancer/chemopreventive agent and is currently used for protection against cancer development/recurrence,with minimal side effects.In this study,we performed gene-set enrichment analysis and found that down-regulated genes induced by 4-HPR were associated with FLT3-ITD gene sets.CD34+AML stem/progenitor cells separated from 32 AML samples were treated with 4-HPR.Correlation analysis showed that AML cells with FLT3-ITD genetic alteration were more sensitive to 4-HPR treatment than those without FLT3-ITD.Next,we treated 22 primary AML cells with 4-HPR and found that 4-HPR was more toxic to AML cells with FLT3-ITD.These results indicated that 4-HPR was preferentially cytotoxic to all FLT3-ITD AML cells irrespective of stem/progenitor cells or blast cells.4-HPR-induced reactive oxygen species(ROS)production and NF-kB inhibition might be the reason of 4-HPR selectivity on FLT3 mutated AML cells.展开更多
Summary： Patients with FLT3-ITD^mmutt/NPM1- cytogenetically normal acute myeloid leukemia （CN-AML）, as high-risk molecular group in CN-AML, are associated with a worse prognosis than other CN-AML patients. It is be...Summary： Patients with FLT3-ITD^mmutt/NPM1- cytogenetically normal acute myeloid leukemia （CN-AML）, as high-risk molecular group in CN-AML, are associated with a worse prognosis than other CN-AML patients. It is beneficial to generate xenotransplantation model of FLT3-ITD^mut/NPM1- CN-AML to better understand the pathogenesis and therapeutic strategies of such AML subtype. The purpose of present study was to establish the xenotransplantation model in NOD/SCID mice with FLT3-ITD^mut/NPM1- CN-AML primary cells. The FLT3-ITD^mut/NPM1- CN-AML primary cells from 3 of 7 cases were successfully transplanted into NOD/SCID mice, and human CD45 positive cells were detected in the peripheral blood, spleen and bone marrow of mice by using flow cytometry. Infiltration of human leukemia cells in various organs of mice was observed by using immunohistochemistry. Gene analysis confirmed sustained FLT3/ITD mutation without NPM1 mutation in mice. By performing serial transplantation, it was found that characteristics of the leukemia cells in secondary and tertiary genera- tion models remained unchanged. Moreover, in vivo cytarabine administration could extend survival of NOD/SCID mice, which was consistent with clinical observation. In conclusion, we successfully estab- lished xenotransplantation model of human FLT3-ITD^mut/NPM1- CN-AML in NOD/SCID mice. The model was able to present primary disease and suitable to evaluate the curative effects of new drugs or therapy strategies.展开更多
Acute myeloid leukemia(AML)is an aggressive hematologic malignancy characterized by poor clinical outcomes,frequently exacerbated by mutations in the FMS-like tyrosine kinase 3(FLT3)gene.Although FLT3 inhibitors(FLT3i...Acute myeloid leukemia(AML)is an aggressive hematologic malignancy characterized by poor clinical outcomes,frequently exacerbated by mutations in the FMS-like tyrosine kinase 3(FLT3)gene.Although FLT3 inhibitors(FLT3i)have emerged as promising therapeutic agents,the absence of molecular biomarkers to predict FLT3i response remains a critical limitation in clinical practice.In this study,we performed a comprehensive multi-omics analysis integrating transcriptomic,proteomic,and pharmacogenomic data from the Beat AML cohort,the Cancer Cell Line Encyclopedia(CCLE),and the PXD023201 repository to elucidate the molecular consequences of FLT3 mutations in AML.Our analysis revealed significant differences in RNA and protein expression profiles between FLT3-mutant and wild-type AML cases,with a particularly striking association between FLT3 mutations and immune suppression.We further evaluated the drug sensitivity of FLT3-mutant patients to 3 FDA-approved FLT3i,gilteritinib,midostaurin,and quizartinib,and observed heightened sensitivity in FLT3-mutant cohorts,accompanied by the activation of immune-related pathways in treatment-responsive groups.These findings suggest a potential synergy between FLT3i efficacy and immune activation.Through rigorous bioinformatic analysis,we identified 3 candidate biomarkers:CD36,SASH1,and NIBAN2,associated with FLT3i sensitivity.These biomarkers were consistently upregulated in favorable prognostic subgroups and demonstrated strong correlations with immune activation pathways.The identification of CD36,SASH1,and NIBAN2 as predictive biomarkers offers a novel toolset for stratifying FLT3i response and prognosis.展开更多
The cooccurrence of NPM1,FLT3-ITD,and DNMT3A mutations(i.e.,triple mutation)is related to dismal prognosis in patients with acute myeloid leukemia(AML)receiving chemotherapy alone.In this multicenter retrospective coh...The cooccurrence of NPM1,FLT3-ITD,and DNMT3A mutations(i.e.,triple mutation)is related to dismal prognosis in patients with acute myeloid leukemia(AML)receiving chemotherapy alone.In this multicenter retrospective cohort study,we aimed to identify whether allogeneic hematopoietic stem cell transplantation(allo-HSCT)could overcome the poor prognosis of DNMT3A^(mut)NPM1^(mut)FLT3-ITD^(mut)AML across four transplant centers in China.Fifty-three patients with triple-mutated AML receiving allo-HSCT in complete remission were enrolled.The 1.5-year probabilities of relapse,leukemia-free survival,and overall survival after allo-HSCT were 11.9%,80.3%,and 81.8%,respectively.Multivariate analysis revealed that more than one course of induction chemotherapy and allo-HSCT beyond CR1 were associated with poor survival.To our knowledge,this work is the largest study to explore the up-to-date undefined role of allo-HSCT in patients with triple-mutated AML.Our real-world data suggest that allo-HSCT could overcome the poor prognosis of DNMT3A^(mut)NPM1^(mut)FLT3-ITD^(mut)in AML.展开更多
文摘Objective： To analyze Fms-like tyrosine kinase 3 （FLT3）/internal-tandem duplications （ITD） mutations in various kinds of hematologic malignancy patients. Methods： FLT3/ITD gene mutations were detected by polymerase chain reaction （PCR） in 103 acute myeloid leukemia （AML） cases, 63 acute lymphocytic leukemia （ALL） cases, 53 chronic myelogenous leukemia （CML） cases in chronic phase （CML-CP）, 34 CML cases in blast crisis （CML-BC）, 11 chronic lymphatic leukemia （CLL） cases, 36 myelodysplastic syndrome （MDS） cases, 9 multiple myeloma （MM） cases and 13 non-hodgkin＇s lymphoma （NHL） cases with marrow infiltration. Results： The expressions of FLT3/ITD gene mutations were detected in 22.3% AML cases, in 6.5% CML-BC cases, in 5.6% MDS cases and in 2.6% ALL cases. The two ALL cases with FLT3/ITD mutation were diagnosed as ALL-L2 with morphology and both with myeloid antigen expression, but finally were diagnosed as acute mixed-lineage leukemia after immunology examination. FLT3/ITD gene mutations were not detected in CML-CP, MM, NHL and CLL cases. In the 23 AML patients with FLT3/ITD gene mutation, including 2 of 8 M1 （2.5%）, 8 of 33 M2 （24.2%）, 7 of 24 M3 （29.3%）, 2 of 11 M4 （18.2%）, 3 of 21 M5 （14.3%）, 1 of 5 M6 （20%）, and 0 of 1 M7 cases, and there were no significant differences in the positive rates of FLT3/ITD mutations between the FAB subtypes （P 〉 0.05）. Statistical analyses showed that in AML patients, FLT3/ITD was associated with a higher peripheral blood white cell （WBC） counts [（41.23 ± 32.56） x 109/L vs （11.36 ± 9.89） × 10^9/L （P 〈 0.01 ）], higher percentage of bone marrow blast cells [（72.78 ± 21.79）% vs （51.26 ± 20.78）% （P 〈 0.05）], and higher cumulative relapse rates （63.6% vs 27.7%, P 〈 0.025） than those negative. Conclusion： FLT3/ITD gene mutation mainly occurred in AML patients, and might be a strong prognostic factor which was associated with high peripheral WBC counts, bone marrow blast cell proportion and a increased relapse risk in AML. Detection of FLT3/ITD gene mutation might provide insights to explore a more accurate genotyping of leukemia, differential diagnosis between AML and ALL, subdivide risk level in AML and estimate prognosis of leukemia.
基金partially funded by the National Natural Science Foundation of China(No.81300401)St.Baldrick’s Foundation International Scholar(No.581580)+1 种基金Natural Science Foundation of Guangdong Province(No.2015A030313460)Guangzhou Women and Children’s Medical Center(No.IP-2008-001 and No.GCP-2019-006).
文摘FMS-like tyrosine kinase 3(FLT3)mutation is strongly associated with poor prognosis in acute myeloid leukemia(AML).Though many FLT3 inhibitors have been developed for clinical application with 34%-56%complete remission rate,patients would develop resistance sooner or later after initial response to tyrosine kinase inhibitors(TKIs),such as gilteritinib.And increasing studies have shown that several resistance related mutations of FLT3 emerged during the AML progression.Thus,further investigation is warranted for these FLT3mu,AML patients to achieve a better treatment outcome.4-Hydroxyphenyl retinamide(4-HPR)has been investigated extensively in animal models and clinical trials as an anticancer/chemopreventive agent and is currently used for protection against cancer development/recurrence,with minimal side effects.In this study,we performed gene-set enrichment analysis and found that down-regulated genes induced by 4-HPR were associated with FLT3-ITD gene sets.CD34+AML stem/progenitor cells separated from 32 AML samples were treated with 4-HPR.Correlation analysis showed that AML cells with FLT3-ITD genetic alteration were more sensitive to 4-HPR treatment than those without FLT3-ITD.Next,we treated 22 primary AML cells with 4-HPR and found that 4-HPR was more toxic to AML cells with FLT3-ITD.These results indicated that 4-HPR was preferentially cytotoxic to all FLT3-ITD AML cells irrespective of stem/progenitor cells or blast cells.4-HPR-induced reactive oxygen species(ROS)production and NF-kB inhibition might be the reason of 4-HPR selectivity on FLT3 mutated AML cells.
基金supported by the National Natural Science Foundation of China (No. 81200380)
文摘Summary： Patients with FLT3-ITD^mmutt/NPM1- cytogenetically normal acute myeloid leukemia （CN-AML）, as high-risk molecular group in CN-AML, are associated with a worse prognosis than other CN-AML patients. It is beneficial to generate xenotransplantation model of FLT3-ITD^mut/NPM1- CN-AML to better understand the pathogenesis and therapeutic strategies of such AML subtype. The purpose of present study was to establish the xenotransplantation model in NOD/SCID mice with FLT3-ITD^mut/NPM1- CN-AML primary cells. The FLT3-ITD^mut/NPM1- CN-AML primary cells from 3 of 7 cases were successfully transplanted into NOD/SCID mice, and human CD45 positive cells were detected in the peripheral blood, spleen and bone marrow of mice by using flow cytometry. Infiltration of human leukemia cells in various organs of mice was observed by using immunohistochemistry. Gene analysis confirmed sustained FLT3/ITD mutation without NPM1 mutation in mice. By performing serial transplantation, it was found that characteristics of the leukemia cells in secondary and tertiary genera- tion models remained unchanged. Moreover, in vivo cytarabine administration could extend survival of NOD/SCID mice, which was consistent with clinical observation. In conclusion, we successfully estab- lished xenotransplantation model of human FLT3-ITD^mut/NPM1- CN-AML in NOD/SCID mice. The model was able to present primary disease and suitable to evaluate the curative effects of new drugs or therapy strategies.
基金supported by grants from the CAMS Innovation Fund for Medical Science(2023-I2M-3-014 to H.W.,2023-I2M-2-007 to H.W.,2021-I2M-1-073 to H.W.)the National Natural Science Foundation of China(82270236 to H.W.,82341080 to H.W.,82400271 to M.N.,82273217 to L.S.)+2 种基金the Fundamental Research Funds for the Central Universities,Peking Union Medical College(3332024200 to M.N.)the distinguished Young Scholars of Tianjin(22JCJQJC00090 to H.W.)the Tianjin Municipal Science and Technology Commission Grant(22JCQNJC00040 to L.S.).
文摘Acute myeloid leukemia(AML)is an aggressive hematologic malignancy characterized by poor clinical outcomes,frequently exacerbated by mutations in the FMS-like tyrosine kinase 3(FLT3)gene.Although FLT3 inhibitors(FLT3i)have emerged as promising therapeutic agents,the absence of molecular biomarkers to predict FLT3i response remains a critical limitation in clinical practice.In this study,we performed a comprehensive multi-omics analysis integrating transcriptomic,proteomic,and pharmacogenomic data from the Beat AML cohort,the Cancer Cell Line Encyclopedia(CCLE),and the PXD023201 repository to elucidate the molecular consequences of FLT3 mutations in AML.Our analysis revealed significant differences in RNA and protein expression profiles between FLT3-mutant and wild-type AML cases,with a particularly striking association between FLT3 mutations and immune suppression.We further evaluated the drug sensitivity of FLT3-mutant patients to 3 FDA-approved FLT3i,gilteritinib,midostaurin,and quizartinib,and observed heightened sensitivity in FLT3-mutant cohorts,accompanied by the activation of immune-related pathways in treatment-responsive groups.These findings suggest a potential synergy between FLT3i efficacy and immune activation.Through rigorous bioinformatic analysis,we identified 3 candidate biomarkers:CD36,SASH1,and NIBAN2,associated with FLT3i sensitivity.These biomarkers were consistently upregulated in favorable prognostic subgroups and demonstrated strong correlations with immune activation pathways.The identification of CD36,SASH1,and NIBAN2 as predictive biomarkers offers a novel toolset for stratifying FLT3i response and prognosis.
基金supported by the National Key Research and Development Program of China(Nos.2022YFC2502600 and 2022YFC2502606)the National Natural Science Foundation of China(Nos.82170206 and 82170208)+3 种基金CAMS Innovation Fund for Medical Sciences(Nos.2019-I2M-5-034 and 2022-I2M-C&T-B-121)Peking University People’s Hospital Research and Development Funds(No.RZ2022-02)Tongzhou District Distinguished Young Scholars(No.JCQN2023009)Shanghai Municipal Health Commission Project of Disciplines of Excellence(No.20234Z0002)。
文摘The cooccurrence of NPM1,FLT3-ITD,and DNMT3A mutations(i.e.,triple mutation)is related to dismal prognosis in patients with acute myeloid leukemia(AML)receiving chemotherapy alone.In this multicenter retrospective cohort study,we aimed to identify whether allogeneic hematopoietic stem cell transplantation(allo-HSCT)could overcome the poor prognosis of DNMT3A^(mut)NPM1^(mut)FLT3-ITD^(mut)AML across four transplant centers in China.Fifty-three patients with triple-mutated AML receiving allo-HSCT in complete remission were enrolled.The 1.5-year probabilities of relapse,leukemia-free survival,and overall survival after allo-HSCT were 11.9%,80.3%,and 81.8%,respectively.Multivariate analysis revealed that more than one course of induction chemotherapy and allo-HSCT beyond CR1 were associated with poor survival.To our knowledge,this work is the largest study to explore the up-to-date undefined role of allo-HSCT in patients with triple-mutated AML.Our real-world data suggest that allo-HSCT could overcome the poor prognosis of DNMT3A^(mut)NPM1^(mut)FLT3-ITD^(mut)in AML.
