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
BACKGROUND Congenital lymphangiectasia is a rare disease characterized by dilated interstitial lymphatic vessels and cystic expansion of the lymphatic vessels.Congenital lymphangiectasia can affect various organ syste...BACKGROUND Congenital lymphangiectasia is a rare disease characterized by dilated interstitial lymphatic vessels and cystic expansion of the lymphatic vessels.Congenital lymphangiectasia can affect various organ systems;however,it frequently occurs in the lungs accompanied with unexplained pleural effusion.Further,it might not be diagnosed during prenatal examination owing to the absence of pronounced abnormalities.However,after birth the newborn rapidly develops respiratory distress that quickly deteriorates.Genetic variations in proteins controlling the development of lymphatic vessels contribute to the pathophysiology of this disease.We report a rare case of heterozygous mutation of ADAMTS3 and FLT4 genes,which have not been reported previously.CASE SUMMARY We analysed the case of a neonate who had presented with only pleural effusion at a late gestational age and eventually died due to its inability to establish spontaneous breathing after birth.An autopsy revealed lymphangiectasia of the organ systems.Further,whole exome sequencing revealed heterozygous mutations of the lymphangiogenesis-controlling genes,ADAMTS3 and FLT4,and Sanger verification revealed similar lesions in the mother with no symptoms.CONCLUSION Considering the presented case,obstetricians should observe unexplained foetal pleural effusion,and perform pathology analysis and whole exome sequencing for a conclusive diagnosis and prompt treatment.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
目的探讨FLT3内部串联重复(internal tandem duplication,ITD)与C-Kit基因突变与急性白血病突变预后相关性。方法利用PCR扩增技术结合DNA测序技术分析33例急性髓系白血病(AML),13例急性淋巴细胞白血病(ALL),急性混合细胞白血病6例患者...目的探讨FLT3内部串联重复(internal tandem duplication,ITD)与C-Kit基因突变与急性白血病突变预后相关性。方法利用PCR扩增技术结合DNA测序技术分析33例急性髓系白血病(AML),13例急性淋巴细胞白血病(ALL),急性混合细胞白血病6例患者骨髓单个核细胞中FLT3/ITD基因与C-Kit基因突变率,分析FLT3/ITD基因与C-Kit基因突变阳性患者的表达水平及疗效。结果急性髓系白血病(AML)患者的FLT3/ITD基因及C-Kit基因平均表达水平高于急性淋巴细胞白血病患者(P<0.05)。33例初治急性白血病FLT3/ITD阳性及C-Kit阳性的患者治疗后缓解率低于阴性患者。结论 FLT3/ITD及C-Kit基因是急性白血病中常见的基因突变类型,均属于III型受体酪氨酸激酶家族成员,与白血病的发生及预后有相关性。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
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The Wu Jieping Medical Foundation Clinical Research Special Grant Fund in China,No.320.6750.2022-15-9.
文摘BACKGROUND Congenital lymphangiectasia is a rare disease characterized by dilated interstitial lymphatic vessels and cystic expansion of the lymphatic vessels.Congenital lymphangiectasia can affect various organ systems;however,it frequently occurs in the lungs accompanied with unexplained pleural effusion.Further,it might not be diagnosed during prenatal examination owing to the absence of pronounced abnormalities.However,after birth the newborn rapidly develops respiratory distress that quickly deteriorates.Genetic variations in proteins controlling the development of lymphatic vessels contribute to the pathophysiology of this disease.We report a rare case of heterozygous mutation of ADAMTS3 and FLT4 genes,which have not been reported previously.CASE SUMMARY We analysed the case of a neonate who had presented with only pleural effusion at a late gestational age and eventually died due to its inability to establish spontaneous breathing after birth.An autopsy revealed lymphangiectasia of the organ systems.Further,whole exome sequencing revealed heterozygous mutations of the lymphangiogenesis-controlling genes,ADAMTS3 and FLT4,and Sanger verification revealed similar lesions in the mother with no symptoms.CONCLUSION Considering the presented case,obstetricians should observe unexplained foetal pleural effusion,and perform pathology analysis and whole exome sequencing for a conclusive diagnosis and prompt treatment.
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.
文摘目的探讨FLT3内部串联重复(internal tandem duplication,ITD)与C-Kit基因突变与急性白血病突变预后相关性。方法利用PCR扩增技术结合DNA测序技术分析33例急性髓系白血病(AML),13例急性淋巴细胞白血病(ALL),急性混合细胞白血病6例患者骨髓单个核细胞中FLT3/ITD基因与C-Kit基因突变率,分析FLT3/ITD基因与C-Kit基因突变阳性患者的表达水平及疗效。结果急性髓系白血病(AML)患者的FLT3/ITD基因及C-Kit基因平均表达水平高于急性淋巴细胞白血病患者(P<0.05)。33例初治急性白血病FLT3/ITD阳性及C-Kit阳性的患者治疗后缓解率低于阴性患者。结论 FLT3/ITD及C-Kit基因是急性白血病中常见的基因突变类型,均属于III型受体酪氨酸激酶家族成员,与白血病的发生及预后有相关性。
基金This work was 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.
