Construction method and application of hematopoietic system-specific bcl11a deletion-induced acute t cell leukemia mouse model

Abstract

The application belongs to the technical field of biological medicine, and relates to a construction method of a mouse model of acute T cell leukemia induced by hematopoietic system specific Bcl11a deletion. Vavl-Cre ; Bcl11a fl / fl The mouse spontaneously develops acute T lymphocytic leukemia (T-ALL); the transgenic mouse is crossed with the Vavl-Cre Bcl11a fl / fl mouse to obtain the hematopoietic system specific Bcl11a deletion Vavl-Cre ; Bcl11a fl / fl mouse. The model mouse spontaneously develops T-ALL without exogenous introduction of a cancer gene, virus infection or chemical induction. The model can stably develop malignant proliferation of thymus-derived T cells, and form a disease phenotype and molecular characteristics highly similar to human T-ALL, and can be used as an important experimental platform for studying the pathogenesis of T-ALL, the disease progression process and carrying out anti-T-ALL drug screening and efficacy evaluation, and can be used for anti-T-ALL candidate drug screening, efficacy evaluation and related target verification research.​

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the construction method and application of a mouse model of acute T-cell leukemia induced by hematopoietic system-specific Bcl11a deletion. Background Technology

[0002] T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive hematologic malignancy originating from thymic T-cell precursors. Its development is typically accompanied by pathological processes such as T-cell developmental arrest, abnormal proliferation, enhanced anti-apoptotic capacity, and clonal expansion. T-ALL patients often exhibit abnormal T-cell proliferation and infiltration in the bone marrow, peripheral blood, thymus, spleen, lymph nodes, and other tissues and organs. Although recent research has made some progress in understanding the molecular mechanisms of T-ALL, its pathogenesis remains incompletely elucidated, and relapse and drug resistance remain important factors affecting patient prognosis. Therefore, establishing animal models that accurately simulate the development and progression of human T-ALL is of great significance for in-depth research into the pathogenesis of T-ALL and for drug screening.

[0003] Currently, human T-ALL commonly exhibits molecular characteristics such as abnormal activation of the Notch1 pathway, abnormal cell cycle regulation, abnormal apoptosis pathway, and abnormal expression of T cell development-related transcription factors. Therefore, an ideal T-ALL animal model should not only exhibit a typical leukemia phenotype but also possess similar pathological features, gene mutation characteristics, transcriptomic characteristics, and tumor cell passage ability to human T-ALL, thereby meeting the needs of pathogenesis research, drug screening, and efficacy evaluation. Currently, animal models used for T-ALL research mainly include the following categories: (1) Oncogene transgene or overexpression model These models are typically established by overexpressing T-ALL-related oncogenes in the mouse hematopoietic or T-cell systems, such as the Notch1 activation model, the Lmo2 transgenic model, and the Tal1 transgenic model. These models can induce T-cell leukemia in mice and can be used to study the role of specific oncogenes in the development of T-ALL. However, these models often rely on the forced expression of exogenous oncogenes and cannot fully reflect the natural occurrence of T-ALL.

[0004] (2) Virus-induced or gene transduction model These models typically involve introducing activated oncogenes such as Notch1, Lmo2, and Tal1 into hematopoietic progenitor cells via retroviruses or lentiviruses, followed by transplantation into recipient mice to induce T-ALL. While this method offers rapid modeling, it relies on artificial gene transduction and transplantation, resulting in differences in the disease's pathogenesis compared to the natural course.

[0005] (3) Leukemia cell transplantation model These models are established by transplanting malignantly transformed T-ALL cells into recipient mice. Based on the donor cell source, they can be further divided into syngeneic transplantation models of mouse-derived T-ALL cells, xenograft models derived from human T-ALL cell lines (cell line-derived xenograft, CDX), and patient-derived xenograft models (PDX). These models are commonly used for drug efficacy evaluation and drug sensitivity testing, but they typically reflect the expansion and infiltration of leukemia cells in the recipient body, rather than simulating the early process of malignant transformation of T-ALL from normal thymic progenitor cells. Furthermore, PDX models rely on immunodeficient receptors and cannot fully simulate the thymic microenvironment and the natural development of T-ALL in mice.

[0006] In summary, while existing T-ALL animal models have played an important role in T-ALL mechanism research and drug evaluation, they still suffer from drawbacks such as strong dependence on exogenous oncogenes or the transplantation process, insufficient simulation of the natural pathogenesis process, and difficulty in being stably used for pathogenesis research and drug screening. Therefore, there is still a need to establish a novel animal model that can spontaneously generate T-ALL, has a stable phenotype, can highly simulate the key characteristics of human T-ALL, and can be used for pathogenesis research and drug screening. Summary of the Invention

[0007] To address the problems existing in the prior art, this invention aims to provide a method for constructing and applying a mouse model of acute T-cell leukemia induced by hematopoietic system-specific Bcl11a deletion. This invention employs a Cre-LoxP conditional gene knockout strategy to construct a mouse model of hematopoietic system-specific Bcl11a deletion (… Vav1-Cre ; Bcl11a fl / fl This study aims to establish an animal model of T-cell acute lymphoblastic leukemia (T-ALL) that can spontaneously develop without the introduction of exogenous oncogenes, viral infection, or chemical induction. This model stably develops malignant proliferation of thymic-derived T cells and exhibits a disease phenotype and molecular characteristics highly similar to human T-ALL. This model can stably develop T-ALL without the introduction of exogenous oncogenes or chemical induction, possesses highly similar pathological, gene mutation, and transcriptomic characteristics to human T-ALL, and has the ability to be serially transplanted. It can serve as an important experimental platform for studying the pathogenesis and progression of T-ALL, as well as for screening and evaluating the efficacy of anti-T-ALL drugs.

[0008] To achieve the above-mentioned objectives, the present invention adopts the following technical solution: The first aspect of the present invention provides a method for constructing a mouse model of acute T-cell leukemia induced by hematopoietic system-specific Bcl11a deletion, using a Cre-LoxP conditional gene knockout strategy to construct a mouse model of hematopoietic system-specific Bcl11a deletion. Vav1-Cre ; Bcl11a fl / fl Mice were used to induce spontaneous development of acute T-lymphoblastic leukemia (T-ALL); the construction method included the following steps: Will Vav1-Cre Transgenic mice and Bcl11a fl / fl Mice were hybridized to obtain hematopoietic system-specific Bcl11a deletions. Vav1-Cre ; Bcl11a fl / fl Mice; The Vav1-Cre Transgenic mice are carriers Vav1-Cre Transgenic mice, the Bcl11a fl / fl Mice are carriers Bcl11a fl / fl Mice with conditional knockout of alleles.

[0009] Furthermore, the construction method also includes the step of identifying the genotype of the hybrid offspring, and obtaining mice with the target genotype through genotype identification.

[0010] This invention employs the Cre-LoxP conditional gene knockout strategy to construct a hematopoietic system-specific Bcl11a deletion mouse model. The Cre-LoxP system is a widely used conditional gene knockout technology that uses Cre recombinase to recognize the LoxP sequence and mediate the deletion of the DNA fragment within it, achieving tissue-specific inactivation of the target gene. Vav1-Cre The transgenic mice carry Cre recombinase expressed under the drive of the Vav1 promoter. Vav1 is a signal transduction protein specifically expressed in hematopoietic stem cells and their progeny hematopoietic cells. Therefore, Cre recombinase can be stably expressed in hematopoietic cells, enabling specific recombination of the target gene in the hematopoietic system.

[0011] Bcl11a (B-cell leukemia / lymphoma 11A) is a zinc finger transcription factor that plays an important regulatory role in the development of hematopoietic stem cells, lymphocytes, and thymic progenitor cells. Bcl11a fl / flThe mice were conditional knockout mice, with LoxP sites (floxed alleles) inserted flanking the key exons of the Bcl11a gene. When Cre recombinase was expressed, it mediated specific recombination of the DNA sequence between the two LoxP sites, thereby inactivating the Bcl11a gene.

[0012] This invention will... Vav1-Cre Transgenic mice and Bcl11a fl / fl Mice were hybridized to obtain blood system-specific Bcl11a-deficient mice ( Vav1-Cre;Bcl11a fl / fl In this model, Bcl11a is specifically lost throughout the hematopoietic system, starting from the hematopoietic stem cell stage, including hematopoietic stem cells, common lymphoid progenitor cells, and thymic progenitor cells.

[0013] A second aspect of this invention provides a mouse model of acute T-cell leukemia obtained by the above-described construction method. The Bcl11a gene is specifically knocked out and inactivated in mouse hematopoietic cells, and the animal model spontaneously develops acute T-lymphoblastic leukemia without the need for exogenous oncogene introduction, viral infection, or chemical induction.

[0014] Furthermore, the mouse model of acute T-cell leukemia was able to stably develop malignant proliferation of thymus-derived T cells and form a disease phenotype and molecular characteristics highly similar to those of human T-ALL.

[0015] Furthermore, the aforementioned acute T-cell leukemia mouse model possesses one or more of the following characteristics: (1) Enlargement of the thymus, spleen, and lymph nodes; (2) Leukemia cells extensively infiltrate the liver, kidneys, bone marrow, and peripheral lymphoid tissues; (3) Tumor cells carry Notch1 mutations, accompanied by activation of the Notch signaling pathway, increased expression of Myc and Hes1 genes, and cell cycle and mTOR signaling pathway activation characteristics. (4) Infiltration of immature thymus-derived T cells in peripheral blood; (5) Tumor cells can be continuously transplanted into recipient mice.

[0016] The third aspect of this invention provides the application of the above-mentioned acute T-cell leukemia mouse model in the preparation of drugs for screening and efficacy evaluation against acute T-lymphoblastic leukemia.

[0017] The animal model established in this invention can stably develop T-ALL without the introduction of exogenous oncogenes, viral infection, or chemical induction, and can effectively simulate the natural occurrence and development of the disease. Furthermore, this model exhibits pathological, gene mutation, and transcriptomic characteristics highly similar to human T-ALL, and possesses stable continuous transplantation capability.

[0018] This invention further provides the application of this model in the study of T-ALL pathogenesis, leukemia initiation cells, drug screening, efficacy evaluation, and the development of novel anti-T-ALL treatment strategies, thereby providing a reliable experimental platform for basic and clinical translational research on T-ALL.

[0019] The invention constructs Vav1-Cre;Bcl11a fl / fl Mouse models can effectively simulate the natural occurrence and progression of human T-ALL, serving as an important experimental platform for studying the pathogenesis of T-ALL. They can also be used for screening anti-T-ALL candidate drugs, evaluating efficacy, and validating related targets.

[0020] Compared with existing T-ALL animal models, the hematopoietic system-specific Bcl11a deletion-induced acute T-cell leukemia mouse model established in this invention has the following beneficial effects: (1) T-ALL can occur spontaneously without exogenous induction. This invention is the first to discover that blood system-specific Bcl11a deficiency can induce spontaneous acute T-lymphoblastic leukemia (T-ALL) in mice. The invention constructs... Vav1-Cre ; Bcl11a fl / fl Mice can stably develop T-ALL without the introduction of exogenous oncogenes, viral infection, or chemical induction, which is closer to the natural occurrence of the disease.

[0021] (2) It can better simulate the key pathological features of human T-ALL. The model of this invention can exhibit typical pathological features such as significant enlargement of the thymus, spleen and lymph nodes, and extensive infiltration of leukemia cells into the liver, kidneys, bone marrow and peripheral lymphoid tissues, which are highly similar to human T-ALL.

[0022] (3) It has molecular characteristics that are highly similar to those of human T-ALL. The model of this invention detects high-frequency Notch1 mutations, accompanied by activation of the Notch signaling pathway, abnormal expression of classic T-ALL-related genes such as Myc and Hes1, and features such as cell cycle and mTOR signaling pathway activation, which can better simulate the molecular pathological process of human T-ALL.

[0023] (4) The disease is stable and highly recurrent. The model of this invention has a high and stable incidence rate and good consistency in disease phenotype, which can provide stable and reliable experimental materials for the study of T-ALL pathogenesis and improve the reproducibility of experimental results.

[0024] (5) Has the ability to perform continuous transplantation The leukemia cells derived from the model of this invention can be stably expanded and continuously passaged in recipient mice, maintaining the original disease characteristics, facilitating the establishment of a standardized experimental system, and suitable for long-term mechanism research and drug evaluation.

[0025] (6) Applicable to the study of the pathogenesis of T-ALL The model of this invention can simulate the entire process of transformation from normal thymic progenitor cells to leukemia cells, and can be used to study the mechanisms of T-ALL occurrence, development, invasion and relapse, providing experimental evidence for elucidating the pathogenesis of T-ALL.

[0026] (7) Applicable to the screening and efficacy evaluation of anti-T-ALL drugs. The model of this invention has stable disease pathogenesis, clear disease progression, and continuous transplantation capability. It can be used for candidate drug screening, efficacy evaluation, combination drug research, and drug resistance mechanism research, thereby improving drug development efficiency and reducing research and development costs.

[0027] (8) It has broad application prospects This invention not only provides a spontaneous T-ALL animal model, but also establishes an experimental platform suitable for studying the pathogenesis of T-ALL, drug screening, and efficacy evaluation. It can be used not only for basic T-ALL research, but also for targeted drug development, treatment optimization, and evaluation of personalized treatment strategies, demonstrating high scientific and practical value. Attached Figure Description

[0028] Figure 1 This is a schematic diagram of the animal model construction of the present invention.

[0029] Figure 2 This is a graph showing the phenotypic identification results of the animal model of this invention. Specifically: a. Animal model survival curve; b. Observation of thymus, spleen, and lymph node tissues in the animal model; cd. Results of bone marrow albinism and erythrocyte reduction in the animal model; e. Analysis results of bone marrow infiltration in the kidneys and liver of the animal model; f. Results of thymic progenitor cells infiltrating into the peripheral blood of T-ALL mice; gh. Analysis results of T-ALL serial transplantation.

[0030] Figure 3This is a diagram showing the molecular characterization results of the animal model of this invention. Specifically: a. Schematic diagram of Notch1 signaling pathway mutations; b. Notch1 structural diagram: EGF-like domain, HD, TAD, and PEST domains, with each arrow representing a mutation and each color representing a sample, for a total of 10 samples; c. T-ALL transcriptome differential analysis results; d. Cell cycle and mTOR signal GSEA enrichment analysis results. Detailed Implementation

[0031] To better understand the present invention, it is now further described with reference to the following embodiments and accompanying drawings. The embodiments are for illustrative purposes only and do not limit the invention in any way. In the embodiments, all original reagents and materials are commercially available, and experimental methods not specifically specified are conventional methods and conditions well known in the art, or according to the conditions recommended by the instrument manufacturer.

[0032] This invention provides an animal model of spontaneous acute T-cell lymphoblastic leukemia (T-ALL) constructed based on Bcl11a conditional deletion and its application.

[0033] This invention employs a Cre-LoxP conditional gene knockout strategy to construct hematopoietic system-specific Bcl11a-deficient mice (… Vav1-Cre ; Bcl11a fl / fl We established an animal model of T-ALL that can spontaneously develop without the introduction of exogenous oncogenes, viral infection, or chemical induction. This model can stably develop malignant proliferation of thymic T cells and form a disease phenotype and molecular characteristics highly similar to human T-ALL.

[0034] The invention constructs Vav1-Cre;Bcl11a fl / fl Mouse models can effectively simulate the natural occurrence and progression of human T-ALL, serving as an important experimental platform for studying the pathogenesis of T-ALL. They can also be used for screening anti-T-ALL candidate drugs, evaluating efficacy, and validating related targets.

[0035] The following examples use Vav1-Cre The transgenic mice were purchased from Jackson Labatory. Bcl11a fl / fl Mice: These are conditional knockout mice, with LoxP sites (floxed alleles) inserted on both sides of the key exon of the Bcl11a gene. Bcl11a fl / flThe mice used were the same mice used in previous studies in our laboratory (Reference: A highly efficient recombineering-based method for generating conditional knockout mutations; Bcl11a is essential for lymphoid development and negatively regulates p53).

[0036] Experimental Example 1: Construction of a hematopoietic system-specific Bcl11a deletion animal model This invention employs the Cre-LoxP conditional gene knockout strategy to construct a hematopoietic system-specific Bcl11a deletion mouse model. The Cre-LoxP system is a widely used conditional gene knockout technology that uses Cre recombinase to recognize the LoxP sequence and mediate the deletion of the DNA fragment within it, achieving tissue-specific inactivation of the target gene. Vav1-Cre transgenic mice carry Cre recombinase expressed driven by the Vav1 promoter. Vav1 is a signal transduction protein specifically expressed in hematopoietic stem cells and their progeny hematopoietic cells; therefore, the Cre recombinase can be stably expressed in hematopoietic cells, achieving specific recombination of the target gene within the hematopoietic system.

[0037] Bcl11a (B-cell leukemia / lymphoma 11A) is a zinc finger transcription factor that plays an important regulatory role in the development of hematopoietic stem cells, lymphocytes, and thymic progenitor cells. Bcl11a fl / fl The mice were conditional knockout mice, with LoxP sites (floxed alleles) inserted flanking the key exons of the Bcl11a gene. When Cre recombinase was expressed, it mediated specific recombination of the DNA sequence between the two LoxP sites, thereby inactivating the Bcl11a gene.

[0038] This embodiment will... Vav1-Cre Transgenic mice and Bcl11a fl / fl Mice were hybridized, and mice with the target genotype were obtained through genotyping. These mice were blood system-specific Bcl11a deletion mice. Vav1-Cre;Bcl11a fl / fl In this model, Bcl11a is specifically lost throughout the hematopoietic system, starting from the hematopoietic stem cell stage, including hematopoietic stem cells, common lymphoid progenitor cells, and thymic progenitor cells.

[0039] As attached Figure 1 As shown, the top is Vav1-CreA transgenic mouse model was developed, employing a tissue-specific Cre recombinase expression system integrated into the intron region of the Commd10 gene on mouse chromosome 18. The 5' end indicates the direction of DNA transcription initiation; subsequently, the Vav promoter drives the transcriptional activation of downstream genes in various hematopoietic lineages; downstream of the promoter is the Cre coding sequence; and the 3' end indicates the direction of transcription termination. This model achieves restricted expression of Cre recombinase in hematopoietic cells through a tissue-specific promoter, providing a source of recombinase for subsequent conditional gene knockout. The image below shows a Bcl11a conditional allele mouse model: the 5' end indicates the upstream direction of the gene; downstream of it is the forward loxP site, marked with a black triangle, which is a 34 bp palindromic recombination sequence specifically recognized by Cre recombinase; downstream of the loxP site is exon 4 of the Bcl11a gene, which encodes a key functional domain of the protein, flanked by loxP sites in the same direction to form a fluxed conformation; downstream of Exon 4 is the frt site, marked with a blue triangle, which is the Flp recombinase recognition sequence used to mediate the deletion of the selection marker; downstream of the frt site is the second loxP site in the same direction, aligned with the upstream site; the 3' end indicates the downstream direction of the gene.

[0040] The core design of this model lies in the anchoring of Exon 4 at two co-directional loxP sites. In the presence of Cre recombinase, these sites catalyze site-specific recombination, leading to the functional inactivation of the Bcl11a gene by excision of the Exon 4 fragment. In tissues without Cre expression, the normal wild-type function is maintained. This strategy successfully constructed a hematopoietic system-specific Bcl11a deletion animal model.

[0041] Example 2: Phenotypic Identification of Animal Models To evaluate the disease phenotype of the constructed model, the model obtained in Example 1 was analyzed. Vav1-Cre;Bcl11a fl / fl Mice were followed up for a long period of time. (See attached image) Figure 2 As shown, this model mouse can spontaneously develop acute T-lymphoblastic leukemia without the introduction of exogenous oncogenes, viral infection, or chemical induction.

[0042] The results showed that Vav1-Cre;Bcl11a fl / fl From 3-4 months of age, the model mice gradually exhibited clinical signs such as decreased activity, rough coat, and kyphosis. As the disease progressed, significant enlargement of the thymus, spleen, and peripheral lymph nodes appeared. Further testing revealed that leukemia cells extensively infiltrated tissues such as the liver, kidneys, and bone marrow, with significant albinism observed in the bone marrow. Simultaneously, abnormal infiltration of immature T cells derived from the thymus was detected in the peripheral blood.

[0043] Furthermore, separation in the late T-ALL stage Vav1-Cre;Bcl11afl / fl as well as Vav1-Cre;Bcl11a fl / fl Mouse bone marrow cells (10 6 / NSG receptors) were transplanted into untreated NSG mice, and the survival status of the recipient mice was assessed after transplantation. When the recipient mice that underwent the first transplantation developed late-stage T-ALL symptoms such as rough fur, arched back, and slow movement, bone marrow cells from the first transplantation recipient mice were isolated and... Vav1-Cre;Bcl11a fl / fl Mouse bone marrow cells (10 6 The NSG receptor was transplanted into untreated NSG mice, and the survival status of the recipient mice was assessed after transplantation. The results showed that... Vav1-Cre;Bcl11a fl / fl T-ALL cells can be transplanted into NSG recipient mice for two consecutive generations, as shown in the attached... Figure 2 As shown in g and h, it can be seen that tumor cells derived from this model can stably reconstruct the leukemia phenotype after transplantation into recipient mice, demonstrating continuous transplantation capability and good disease passage stability.

[0044] Example 3: Molecular Characterization of Animal Models To evaluate the similarity between this model and human T-ALL, whole exome sequencing (WES) and RNA-seq sequencing analyses were performed on tumor cells from the model mice.

[0045] As attached Figure 3 As shown, the results indicated that all tested samples carried Notch1 mutations, which were mainly enriched in the HD, TAD, and PEST domains. The expression of classic T-ALL driver genes such as Notch1, Myc, Hes1, and Dtx1 was increased; the cell cycle and mTOR pathway were significantly activated.

[0046] The above results indicate that the mouse model constructed in this invention is highly similar to human T-ALL in terms of genetic characteristics, transcriptomic characteristics, and biological behavior.

[0047] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A method for constructing a mouse model of acute T-cell leukemia induced by hematopoietic system-specific Bcl11a deficiency, characterized in that, A hematopoietic system-specific Bcl11a deletion gene was constructed using the Cre-LoxP conditional gene knockout strategy. Vav1-Cre ; Bcl11a fl / fl Mice were used to induce spontaneous development of acute T-lymphoblastic leukemia (T-ALL); the construction method included the following steps: Will Vav1-Cre Transgenic mice and Bcl11a fl / fl Mice were hybridized to obtain hematopoietic system-specific Bcl11a deletions. Vav1-Cre ; Bcl11a fl / fl Mice; The Vav1-Cre Transgenic mice are carriers Vav1-Cre Transgenic mice, the Bcl11a fl / fl Mice are carriers Bcl11a fl / fl Mice with conditional knockout of alleles.

2. The method for constructing an acute T-cell leukemia mouse model according to claim 1, characterized in that, The construction method further includes: a step of genotyping the hybrid offspring, and obtaining mice with the target genotype through genotyping.

3. The method for constructing an acute T-cell leukemia mouse model according to claim 1, characterized in that, The hematopoietic system includes hematopoietic stem cells, common lymphoid progenitor cells, and thymic progenitor cells.

4. The method for constructing an acute T-cell leukemia mouse model according to claim 1, characterized in that, The Vav1-Cre The transgenic mice carry Cre recombinase expressed under the drive of the Vav1 promoter.

5. The method for constructing an acute T-cell leukemia mouse model according to claim 1, characterized in that, The Bcl11a fl / fl LoxP sites were inserted flanking the key exon of the mouse Bcl11a gene.

6. An acute T-cell leukemia mouse model obtained by the construction method according to any one of claims 1-5.

7. The mouse model of acute T-cell leukemia according to claim 6, characterized in that, The Bcl11a gene in the mouse hematopoietic system cells is specifically knocked out and inactivated, and the animal model can spontaneously develop acute T-lymphoblastic leukemia without the introduction of exogenous oncogenes, viral infection or chemical induction.

8. The mouse model of acute T-cell leukemia according to claim 6, characterized in that, The model can stably induce malignant proliferation of thymic-derived T cells and form a disease phenotype and molecular characteristics highly similar to human T-ALL.

9. The mouse model of acute T-cell leukemia according to claim 8, characterized in that, The animal model has one or more of the following characteristics: (1) Enlargement of the thymus, spleen, and lymph nodes; (2) Leukemia cells extensively infiltrate the liver, kidneys, bone marrow, and peripheral lymphoid tissues; (3) Tumor cells carry Notch1 mutations, accompanied by activation of the Notch signaling pathway, increased expression of Myc and Hes1 genes, and cell cycle and mTOR signaling pathway activation characteristics. (4) Infiltration of immature thymus-derived T cells in peripheral blood; (5) Tumor cells can be continuously transplanted into recipient mice.

10. The use of the acute T-cell leukemia mouse model according to any one of claims 7-9 in the preparation of drugs for screening and efficacy evaluation against acute T-lymphoblastic leukemia.

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