碩士 / 國立臺灣大學 / 醫學檢驗暨生物技術學研究所 / 106 / In acute myeloid leukemia（AML），a hematologic malignancy, there is an urgent need for molecular understanding as well as targeted therapy. Recently, zebrafish has become an important model organism for the study of AML and other blood diseases, due to many advantages of zebrafish, such as short reproductive cycle, a large number of offspring, transparently and observable embryos, easy to breed and grow rapidly. Like higher vertebrates, zebrafish have two major waves of hematopoiesis, the primitive and definitive waves. The hematopoietic genetic system of zebrafish is highly conserved with humans in evolution. Various important human hematopoietic regulatory transcription factors are conservative.
In previous studies, seven patients, who suffer from acute myeloid leukemia (AML), had the same TALDO1-DSN1 fusion gene. Human TALDO1 encodes an enzyme, transaldolase, which is a crucial regulator in pentose phosphate pathway. In previous studies, the abnormality of TALDO1 was associated with the development of different autoimmune diseases and malignancies. Human DSN1 encodes kinetochore proteins which plays an irreplaceable role in cell division. In breast cancer and colorectal cancer, DSN1 is related to the occurrence and metastasis of cancer.
In this study, we use the transgenic zebrafish line, Tg (pTolCG-spi1: TALDO1-DSN1-PA/CG), to display that the human TALDO1-DSN1 fusion gene driven by spi1 (a myeloid-specific promoter) can lead to abnormal hematopoiesis during its life cycle.
We screened transgenic zebrafish for genetically stable transgenes. For the embryo less than 5 days-post-fertilization (dpf), we detected the expression of specific transcription factors in hematopoiesis by qRT-PCR. The result demonstrated that various genes expressed abnormal. Interestingly, gata1, the erythropoiesis transcription factor, surged at several points in time, as well as the granulocyte marker mpo reduced significantly, and subsequently, we use the whole-mount in situ hybridization technique for double confirmation.
For adult fishes aged from 6 to 12 months, peripheral blood and kidney marrow were taken, and followed by made for blood smears. We taken the blood morphological examination by cell typing and counting. The results displayed that in some transgenic adult zebrafish, immature myelo/monocytes appeared in the peripheral blood. In kidney marrow, the percentage of blast cells and myelo/monocytes increased abnormally.
| Identifer | oai:union.ndltd.org:TW/106NTU05108013 |
| Date | January 2018 |
| Creators | Yan Huang, 黃焱 |
| Contributors | 林亮音 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 62 |
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