MYELOID-SPECIFIC EXPRESSION OF SOX4 INDUCES EXPANDED MYELOPOIESIS WITH RETARDED CELL DIFFERENTIATION - A NOVEL ZEBRAFISH LEUKEMIA MODEL
(Abstract release date: 05/19/16)
EHA Library. Lu J. 06/09/16; 134526; PB1626
Dr. Jeng-Wei Lu
Contributions
Contributions
Abstract
Abstract: PB1626
Type: Publication Only
Background
SOX4 belongs to a family of Sox (SRY-related HMG-box) transcription factors, including Sox4, Sox11, and Sox12. Abnormal expression of SOX4 is related to malignant transformation and cancer metastasis. Overexpression of SOX4 is associated with clonal dominance of hematopoietic stem cells, repopulation advantage of various stem/progenitor cells, block in differentiation of myeloid progenitor cells, and induction of myeloid leukemia. Recently, the expression of SOX4, as a direct target of C/EBPα, is reported to inversely correlate with C/EBPα activity. Downregulation of SOX4 attenuates self-renewal capability of leukemic cells and restored their normal differentiation process. SOX4 overexpression resulting from C/EBPα inactivation contributes to the development of leukemia with a distinct leukemia-initiating cell (LIC) phenotype. In addition, several fusion proteins resulting from chromosomal aberration, such as MOZ-TIF2, AML1-ETO, UNP98-HOXA9, were closely linked to SOX4 upregulation, further suggesting the significance of SOX4 in leukemogenesis. Zebrafish is a popular animal model in biomedical researches. Here, we add to its strength for leukemia research by providing a novel transgenic SOX4 zebrafish model.
Aims
To establish SOX4 transgenic zebrafish research model.
Methods
By using Multisite Gateway system® and Tol2 transposon technology, we established a construct including spi1 promoter, human SOX4 gene and EGFP. This construct was then injected into zebrafish embryos. After a series of fluorescent selection, stable transgenic SOX4 zebrafish - Tg(CG-spi1-SOX4-EGFP)were generated. The expression of hematopoiesis-related transcription factors were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) or whole-mount in situ hybridization (WISH) technique. The histological pictures of kidney marrow (KM) were examined by light microscope
Results
We found that SOX4 could be detected at 20 hpf (0.2~1.7x103 copies), and then increased gradually during development. At 5 dpf, level of SOX4 was up to 2~10x103 copies. However, there were no significant differences in the expression of hematopoiesis-related transcription factors, such as csf1r, cebpa, l-plastin, mpeg1, mpo, runx1 and c-myb in spite of the expression of SOX4. These results indicated that the SOX4 transgenic zebrafish had normal hematopoiesis in the larval stage. However, in comparison with age-matched AB-wild type fish, the KM of 5-month old transgenic zebrafish Tg(CG-spi1-SOX4-EGFP) had a greater number of myeloid progenitors (9.24±2.75% vs 5.27±0.74%; p<0.05); those of 9-month old had a greater number of myeloid progenitors (18.00±8.78% vs5.73±0.64, p<0.01), less number of mature erythroid cells (19.55±5.98% vs27.93±6.03%, p<0.05) and greater M:E ratio (10.39±7.81 vs 3.72:1±0.53, p<0.05); those of 12-month old had significant difference in all blood components (17.61±8.07% vs 7.00±1.10% in myeloid progenitors; 62.06±7.82% vs42.07±3.18% in myelomonocyte/neutrophil; 4.11±0.72% vs 16.20±1.12% in lymphocytes; 5.44±1.45% vs 10.06±2.55% in immature erythroid cells;10.78±3.40% vs 4.61±0.64% in mature erythroid cells and M:E ratio of15.77±5.34 vs 4.61±0.64, p<0.05 for all above).
Conclusion
These results indicate that SOX4 transgenic zebrafish develops a gradual expansion of myeloid cells, which fail to differentiate, and ultimately transform into myeloid leukemia.
Session topic: E-poster
Keyword(s): Animal model, CCAAT/enhancer binding protein alpha (C/EBPa), Myeloid malignancies, Zebra fish
Type: Publication Only
Background
SOX4 belongs to a family of Sox (SRY-related HMG-box) transcription factors, including Sox4, Sox11, and Sox12. Abnormal expression of SOX4 is related to malignant transformation and cancer metastasis. Overexpression of SOX4 is associated with clonal dominance of hematopoietic stem cells, repopulation advantage of various stem/progenitor cells, block in differentiation of myeloid progenitor cells, and induction of myeloid leukemia. Recently, the expression of SOX4, as a direct target of C/EBPα, is reported to inversely correlate with C/EBPα activity. Downregulation of SOX4 attenuates self-renewal capability of leukemic cells and restored their normal differentiation process. SOX4 overexpression resulting from C/EBPα inactivation contributes to the development of leukemia with a distinct leukemia-initiating cell (LIC) phenotype. In addition, several fusion proteins resulting from chromosomal aberration, such as MOZ-TIF2, AML1-ETO, UNP98-HOXA9, were closely linked to SOX4 upregulation, further suggesting the significance of SOX4 in leukemogenesis. Zebrafish is a popular animal model in biomedical researches. Here, we add to its strength for leukemia research by providing a novel transgenic SOX4 zebrafish model.
Aims
To establish SOX4 transgenic zebrafish research model.
Methods
By using Multisite Gateway system® and Tol2 transposon technology, we established a construct including spi1 promoter, human SOX4 gene and EGFP. This construct was then injected into zebrafish embryos. After a series of fluorescent selection, stable transgenic SOX4 zebrafish - Tg(CG-spi1-SOX4-EGFP)were generated. The expression of hematopoiesis-related transcription factors were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) or whole-mount in situ hybridization (WISH) technique. The histological pictures of kidney marrow (KM) were examined by light microscope
Results
We found that SOX4 could be detected at 20 hpf (0.2~1.7x103 copies), and then increased gradually during development. At 5 dpf, level of SOX4 was up to 2~10x103 copies. However, there were no significant differences in the expression of hematopoiesis-related transcription factors, such as csf1r, cebpa, l-plastin, mpeg1, mpo, runx1 and c-myb in spite of the expression of SOX4. These results indicated that the SOX4 transgenic zebrafish had normal hematopoiesis in the larval stage. However, in comparison with age-matched AB-wild type fish, the KM of 5-month old transgenic zebrafish Tg(CG-spi1-SOX4-EGFP) had a greater number of myeloid progenitors (9.24±2.75% vs 5.27±0.74%; p<0.05); those of 9-month old had a greater number of myeloid progenitors (18.00±8.78% vs5.73±0.64, p<0.01), less number of mature erythroid cells (19.55±5.98% vs27.93±6.03%, p<0.05) and greater M:E ratio (10.39±7.81 vs 3.72:1±0.53, p<0.05); those of 12-month old had significant difference in all blood components (17.61±8.07% vs 7.00±1.10% in myeloid progenitors; 62.06±7.82% vs42.07±3.18% in myelomonocyte/neutrophil; 4.11±0.72% vs 16.20±1.12% in lymphocytes; 5.44±1.45% vs 10.06±2.55% in immature erythroid cells;10.78±3.40% vs 4.61±0.64% in mature erythroid cells and M:E ratio of15.77±5.34 vs 4.61±0.64, p<0.05 for all above).
Conclusion
These results indicate that SOX4 transgenic zebrafish develops a gradual expansion of myeloid cells, which fail to differentiate, and ultimately transform into myeloid leukemia.
Session topic: E-poster
Keyword(s): Animal model, CCAAT/enhancer binding protein alpha (C/EBPa), Myeloid malignancies, Zebra fish
Abstract: PB1626
Type: Publication Only
Background
SOX4 belongs to a family of Sox (SRY-related HMG-box) transcription factors, including Sox4, Sox11, and Sox12. Abnormal expression of SOX4 is related to malignant transformation and cancer metastasis. Overexpression of SOX4 is associated with clonal dominance of hematopoietic stem cells, repopulation advantage of various stem/progenitor cells, block in differentiation of myeloid progenitor cells, and induction of myeloid leukemia. Recently, the expression of SOX4, as a direct target of C/EBPα, is reported to inversely correlate with C/EBPα activity. Downregulation of SOX4 attenuates self-renewal capability of leukemic cells and restored their normal differentiation process. SOX4 overexpression resulting from C/EBPα inactivation contributes to the development of leukemia with a distinct leukemia-initiating cell (LIC) phenotype. In addition, several fusion proteins resulting from chromosomal aberration, such as MOZ-TIF2, AML1-ETO, UNP98-HOXA9, were closely linked to SOX4 upregulation, further suggesting the significance of SOX4 in leukemogenesis. Zebrafish is a popular animal model in biomedical researches. Here, we add to its strength for leukemia research by providing a novel transgenic SOX4 zebrafish model.
Aims
To establish SOX4 transgenic zebrafish research model.
Methods
By using Multisite Gateway system® and Tol2 transposon technology, we established a construct including spi1 promoter, human SOX4 gene and EGFP. This construct was then injected into zebrafish embryos. After a series of fluorescent selection, stable transgenic SOX4 zebrafish - Tg(CG-spi1-SOX4-EGFP)were generated. The expression of hematopoiesis-related transcription factors were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) or whole-mount in situ hybridization (WISH) technique. The histological pictures of kidney marrow (KM) were examined by light microscope
Results
We found that SOX4 could be detected at 20 hpf (0.2~1.7x103 copies), and then increased gradually during development. At 5 dpf, level of SOX4 was up to 2~10x103 copies. However, there were no significant differences in the expression of hematopoiesis-related transcription factors, such as csf1r, cebpa, l-plastin, mpeg1, mpo, runx1 and c-myb in spite of the expression of SOX4. These results indicated that the SOX4 transgenic zebrafish had normal hematopoiesis in the larval stage. However, in comparison with age-matched AB-wild type fish, the KM of 5-month old transgenic zebrafish Tg(CG-spi1-SOX4-EGFP) had a greater number of myeloid progenitors (9.24±2.75% vs 5.27±0.74%; p<0.05); those of 9-month old had a greater number of myeloid progenitors (18.00±8.78% vs5.73±0.64, p<0.01), less number of mature erythroid cells (19.55±5.98% vs27.93±6.03%, p<0.05) and greater M:E ratio (10.39±7.81 vs 3.72:1±0.53, p<0.05); those of 12-month old had significant difference in all blood components (17.61±8.07% vs 7.00±1.10% in myeloid progenitors; 62.06±7.82% vs42.07±3.18% in myelomonocyte/neutrophil; 4.11±0.72% vs 16.20±1.12% in lymphocytes; 5.44±1.45% vs 10.06±2.55% in immature erythroid cells;10.78±3.40% vs 4.61±0.64% in mature erythroid cells and M:E ratio of15.77±5.34 vs 4.61±0.64, p<0.05 for all above).
Conclusion
These results indicate that SOX4 transgenic zebrafish develops a gradual expansion of myeloid cells, which fail to differentiate, and ultimately transform into myeloid leukemia.
Session topic: E-poster
Keyword(s): Animal model, CCAAT/enhancer binding protein alpha (C/EBPa), Myeloid malignancies, Zebra fish
Type: Publication Only
Background
SOX4 belongs to a family of Sox (SRY-related HMG-box) transcription factors, including Sox4, Sox11, and Sox12. Abnormal expression of SOX4 is related to malignant transformation and cancer metastasis. Overexpression of SOX4 is associated with clonal dominance of hematopoietic stem cells, repopulation advantage of various stem/progenitor cells, block in differentiation of myeloid progenitor cells, and induction of myeloid leukemia. Recently, the expression of SOX4, as a direct target of C/EBPα, is reported to inversely correlate with C/EBPα activity. Downregulation of SOX4 attenuates self-renewal capability of leukemic cells and restored their normal differentiation process. SOX4 overexpression resulting from C/EBPα inactivation contributes to the development of leukemia with a distinct leukemia-initiating cell (LIC) phenotype. In addition, several fusion proteins resulting from chromosomal aberration, such as MOZ-TIF2, AML1-ETO, UNP98-HOXA9, were closely linked to SOX4 upregulation, further suggesting the significance of SOX4 in leukemogenesis. Zebrafish is a popular animal model in biomedical researches. Here, we add to its strength for leukemia research by providing a novel transgenic SOX4 zebrafish model.
Aims
To establish SOX4 transgenic zebrafish research model.
Methods
By using Multisite Gateway system® and Tol2 transposon technology, we established a construct including spi1 promoter, human SOX4 gene and EGFP. This construct was then injected into zebrafish embryos. After a series of fluorescent selection, stable transgenic SOX4 zebrafish - Tg(CG-spi1-SOX4-EGFP)were generated. The expression of hematopoiesis-related transcription factors were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) or whole-mount in situ hybridization (WISH) technique. The histological pictures of kidney marrow (KM) were examined by light microscope
Results
We found that SOX4 could be detected at 20 hpf (0.2~1.7x103 copies), and then increased gradually during development. At 5 dpf, level of SOX4 was up to 2~10x103 copies. However, there were no significant differences in the expression of hematopoiesis-related transcription factors, such as csf1r, cebpa, l-plastin, mpeg1, mpo, runx1 and c-myb in spite of the expression of SOX4. These results indicated that the SOX4 transgenic zebrafish had normal hematopoiesis in the larval stage. However, in comparison with age-matched AB-wild type fish, the KM of 5-month old transgenic zebrafish Tg(CG-spi1-SOX4-EGFP) had a greater number of myeloid progenitors (9.24±2.75% vs 5.27±0.74%; p<0.05); those of 9-month old had a greater number of myeloid progenitors (18.00±8.78% vs5.73±0.64, p<0.01), less number of mature erythroid cells (19.55±5.98% vs27.93±6.03%, p<0.05) and greater M:E ratio (10.39±7.81 vs 3.72:1±0.53, p<0.05); those of 12-month old had significant difference in all blood components (17.61±8.07% vs 7.00±1.10% in myeloid progenitors; 62.06±7.82% vs42.07±3.18% in myelomonocyte/neutrophil; 4.11±0.72% vs 16.20±1.12% in lymphocytes; 5.44±1.45% vs 10.06±2.55% in immature erythroid cells;10.78±3.40% vs 4.61±0.64% in mature erythroid cells and M:E ratio of15.77±5.34 vs 4.61±0.64, p<0.05 for all above).
Conclusion
These results indicate that SOX4 transgenic zebrafish develops a gradual expansion of myeloid cells, which fail to differentiate, and ultimately transform into myeloid leukemia.
Session topic: E-poster
Keyword(s): Animal model, CCAAT/enhancer binding protein alpha (C/EBPa), Myeloid malignancies, Zebra fish
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