CN110438158A - A kind of preparation and application of aggressive lymphomas mouse model - Google Patents
A kind of preparation and application of aggressive lymphomas mouse model Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
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Abstract
The invention belongs to translational medicines and new drug development field, and in particular to a kind of preparation and application of aggressive lymphomas mouse model.The invention discloses design, mating, the propagation methods of a completely new aggressive lymphomas mouse model；Disclose the tumorigenic Gender characteristics and life span of aggressive lymphomas mouse model；The phenotype and tissue signature and cell type for disclosing the lymthoma of aggressive lymphomas mouse model are analyzed.The present invention constructs the mouse model of malignant invasion lymthoma for the first time, in the mouse model lymthoma can spontaneous development invade the histoorgans such as brain, it is closely similar with the feature of human malignant's lymthoma, it can be used for the In vivo study of anti-tumor drug and therapy, provide important research model for the preclinical In vivo study of tumour medicine.
Description
Technical field
The invention belongs to translational medicines and new drug development field, and in particular to a kind of system of aggressive lymphomas mouse model
Standby and application.
Background technique
Myc gene is displaced with immunoglobulin gene in mankind's Berkeley lymthoma, causes c-Myc exception table
It reaches, the final Berkeley lymthoma that induces occurs.By genetic engineering method, by the enhancer element of μ gene and Myc gene, that is, E
μ-Myc is integrated into mouse genome, and building obtains E μ-Myc transgenic mice.In the mouse model, Myc is specifically thin in B
Spontaneous lymphoma occurs for unconventionality expression in born of the same parents' atomization, inducing mouse, shows important work of the Myc gene in lymthoma generation
With.
In the whole life cycle of E μ-Myc transgenic mice, semizygote (Hemizygotes) E μ-Myc mouse
Pre-B cell is largely proliferated in marrow.In the semizygote offspring of wild type female mice and semizygote hero mouse, spontaneous pre-B cell
With B cell lymphoma 15-20 week old incidence be 50%.Lymthoma is derived from the single B of the different times broken up in B cell
Lymphoid cell clone.
B cell non-Hodgkin lymphoma (B-NHL) is the main Types of lymthoma, wherein Diffuse Large B Cell lymphoma
(DLBCL) the proportion highest in B-NHL.Clinically, although the gold chemotherapy regimen CHOP/R-CHOP of lymthoma can
To cure 60% DLBCL patient substantially, but still there is 40% DLBCL patient to CHOP/R-CHOP therapeutic scheme without answering
It answers, most of phase after the treatment (4.7-9.0 months), is further detected that lymthoma invades central nervous system in these patients
Unite CNS, invades in case of CNS, and the therapeutic effect of this kind of patient is very undesirable, and subsequent life cycle is only 2-5 months.
But it is not capable of the animal model of fine simulation human patient in the prior art.
Therefore, relevant lymphoma animal models are developed, the recurrence and CNS of studying lymthoma are invaded, and exploitation
Corresponding effective chemotherapy regimen is of great significance, and is the key that treatment malignant lymphoma.
Summary of the invention
In order to overcome the problems of in the prior art, the purpose of the present invention is to provide a kind of aggressive lymphomas is small
The preparation and application of mouse model, the preclinical study that the aggressive lymphomas mouse model can be used for during new drug development.
To achieve the goals above and other related purposes, the present invention adopts the following technical scheme:
The first aspect of the present invention provides a kind of construction method of aggressive lymphomas mouse model, includes the following steps:
By the UTX gene knockout of spontaneous B cell lymphoma mouse, screening obtains aggressive lymphomas mouse model.
In one embodiment, the lymthoma of the aggressive lymphomas mouse model is invaded to organ outside lymph node.
In one embodiment, the lymthoma of the aggressive lymphomas mouse model is invaded to head.
In one embodiment, brain infringement has occurred in the aggressive lymphomas mouse model.
In one embodiment, the lymthoma of the aggressive lymphomas mouse model is invaded to thoracic cavity.
In one embodiment, thymus gland infringement has occurred in the aggressive lymphomas mouse model.
In one embodiment, the lymthoma of the aggressive lymphomas mouse model is invaded to abdominal cavity.
In one embodiment, abdominal cavity infringement has occurred in the aggressive lymphomas mouse model.
In one embodiment, it is raw that a large amount of blood vessels have occurred around the lymthoma of the aggressive lymphomas mouse model
At.
In one embodiment, lymph of the aggressive lymphomas mouse model than spontaneous B cell lymphoma mouse
The blood vessel of tumor becomes apparent from.
In one embodiment, by the UTX gene knockout in spontaneous B cell lymphoma mouse B cell.
In one embodiment, UTX gene is knocked out using the method for genetic engineering.
In one embodiment, spontaneous B cell lymphoma mouse and the mouse of B cell specific deletion UTX gene are handed over
With breeding, screening obtains aggressive lymphomas mouse model from offspring.
In one embodiment, the spontaneous B cell lymphoma mouse is E μ-Myc transgenic mice.
In one embodiment, the construction method of the aggressive lymphomas mouse model, includes the following steps:
(1) wild-type mice of the female UTX gene by gene targeting modification mates with male E μ-Myc transgenic mice,
Obtain the E μ-Myc transgenic mice that UTX gene passes through gene targeting modification；(2) female UTX gene is modified by gene targeting
Wild-type mice mate with male Cre transgenosis tool mouse, obtain UTX gene special knock-out mice in B cell；(3)
It takes in E μ-Myc transgenic mice and step (2) of the male UTX gene obtained in step (1) by gene targeting modification and obtains
The special knock-out mice mating in B cell of female UTX gene, obtain the E μ-Myc that UTX gene specifically knocks out in B cell and turn
DNA murine, screening obtain aggressive lymphomas mouse model.
In another embodiment, the construction method of the aggressive lymphomas mouse model, includes the following steps:
(1) wild-type mice of the female UTX gene by gene targeting modification mates with male Cre transgenosis tool mouse, obtains UTX
Gene special knock-out mice in B cell；(2) male E μ-Myc transgenic mice and step (1) middle female UTX gene obtained
Special knock-out mice mating, obtains the E μ-Myc transgenic mice that UTX gene specifically knocks out in B cell in B cell, sieve
Choosing obtains aggressive lymphomas mouse model.
The second aspect of the present invention provides the aggressive lymphomas mouse model that aforementioned construction method obtains.
The third aspect of the present invention provides the aggressive lymphomas mouse that aforementioned construction method or aforementioned construction method obtain
Model is used to screen the purposes of aggressive lymphomas therapeutic agent.
The fourth aspect of the present invention provides a kind of method for screening aggressive lymphomas therapeutic agent, comprising steps of will wait
It selects substance to be applied to the aggressive lymphomas mouse model that aforementioned construction method obtains, if aggressive lymphomas weakens, can incite somebody to action
The drug candidate is as candidate aggressive lymphomas therapeutic agent.
Compared with prior art, the invention has the following beneficial effects:
The invention discloses design, mating, the propagation methods of a completely new aggressive lymphomas mouse model；It discloses
The tumorigenic Gender characteristics and life span of aggressive lymphomas mouse model；Disclose aggressive lymphomas mouse model
Lymthoma phenotype and tissue signature and cell type analyze.The present invention constructs the mouse of malignant invasion lymthoma for the first time
Model, in the mouse model lymthoma can spontaneous development invade the histoorgans such as brain, very with the feature of human malignant's lymthoma
It is similar, can be used for the In vivo study of anti-tumor drug and therapy, for tumour medicine preclinical In vivo study provide it is important
Research model.
Detailed description of the invention
Figure 1A: the construction method of aggressive lymphomas mouse model.
Figure 1B: another construction method of aggressive lymphomas mouse model.
Fig. 1 C: E μ-Myc transgenic mice of the UTX gene by gene targeting modification is obtained as experiment contrast.
Fig. 2: the tumorigenic Gender characteristics and life span of aggressive lymphomas mouse model.
Fig. 3: the feature of aggressive lymphomas mouse model.
Fig. 4: the feature supplement of aggressive lymphomas mouse model.
Fig. 5: the origin of cell analysis of aggressive lymphomas mouse model.
Specific embodiment
Main inventive of the present invention it is a kind of can spontaneous generation brain invade and the lymphoma animal models of vascularization, the invention
It can be used for the basic research of aggressive lymphomas and the research of the preclinical animal studies during new drug development.
Compared with other lymphoma mouse models, spontaneous lymphoma in genetic engineering mice and transgene mouse model
Development is considered closer to physiology microenvironment, and the development of these lymthomas is under conditions of non-artificial selection pressure.With exempt from
Epidemic disease deficient mice is compared, these genetic engineering lymphoma mouse models are to be immunized normally, clinically relevant with human lymphoma
Property is stronger.Moreover, these lymthomas are in normal biology and immune environment, cell factor in complete immune system
It can more accurately be studied with the function of effector cell.E μ-Myc transgenic mice is that a typical genetic engineering lymthoma is small
Mouse model, the mouse model carry the E μ enhancer sequence and c myc of immunoglobulin gene heavy chain, lead to pre-
Malignant proliferation occurs for B cell and/or B cell.
The discovery of human cancer genome research, mass mutation or deletion occur in kinds cancer for UTX, and these are mutated
It is much Inactivating mutations, therefore, UTX may be a tumor suppressor gene.It is beaten by CD19-Cre tool mouse and by loxP sequence
The UTX mouse of target mates, and has obtained the mouse that B cell specifically knocks out UTX, after further hybridizing with E μ-Myc transgenic mice,
Completely new a genetic engineering and transgene mouse model are obtained, which can be used for studying the lymthoma of UTX missing
The preclinical animal studies research of feature and new drug occurs.
Before further describing the specific embodiments of the present invention, it should be appreciated that protection scope of the present invention is not limited to down
State specific specific embodiment；It is also understood that term used in the embodiment of the present invention is specific specific in order to describe
Embodiment, rather than limiting the scope of protection of the present invention.The test method of actual conditions is not specified in the following example,
Usually according to normal condition, or according to condition proposed by each manufacturer.
When embodiment provides numberical range, it should be appreciated that except non-present invention is otherwise noted, two ends of each numberical range
Any one numerical value can be selected between point and two endpoints.Unless otherwise defined, the present invention used in all technologies and
Scientific term is identical as the normally understood meaning of those skilled in the art of the present technique.Except specific method, equipment used in embodiment,
Outside material, grasp and record of the invention according to those skilled in the art to the prior art can also be used and this
Any method, equipment and the material of the similar or equivalent prior art of method described in inventive embodiments, equipment, material come real
The existing present invention.
Unless otherwise stated, disclosed in this invention experimental method, detection method, preparation method be all made of this technology neck
Molecular biology, biochemistry, chromatin Structure and the analysis of domain routine, analytical chemistry, cell culture, recombinant DNA technology and
The routine techniques of related fields.These technologies have perfect explanation in the prior art, and for details, reference can be made to Sambrook etc.
MOLECULAR CLONING:A LABORATORY MANUAL, Second edition, Cold Spring Harbor
Laboratory Press, 1989and Third edition, 2001；Ausubel etc., CURRENT PROTOCOLS IN
MOLECULAR BIOLOGY, John Wiley&Sons, New York, 1987and periodic updates；the
Series METHODS IN ENZYMOLOGY, Academic Press, San Diego；Wolffe, CHROMATIN
STRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998；METHODS IN
ENZYMOLOGY, Vol.304, Chromatin (P.M.Wassarman and A.P.Wolffe, eds.), Academic
Press, San Diego, 1999；With METHODS IN MOLECULAR BIOLOGY, Vol.119, Chromatin
Protocols (P.B.Becker, ed.) Humana Press, Totowa, 1999 etc..
Embodiment 1
One, material
Spontaneous B cell lymphoma mouse: that is, E μ-Myc transgenic mice, it buys in Jackson Lab, the U.S. (The
Jackson laboratory)。
UTX gene passes through the wild-type mice of gene targeting modification, constructs and derive from the laboratory Chen Degui (biochemistry
With Institute of Cell Biology).
The strain of all mouse occurred in the present invention is C57BL/6 background.All mouse are dynamic according to the experiment of standard
Object method for breeding is raised in no specified microorganisms (SPF) barrier.
All animal feedings follow Shanghai City management of laboratory animal specification and Chinese Academy of Sciences's Shanghai biochemistry and cell
Biological study institute management of laboratory animal rules and regulations.
Two, aggressive lymphomas mouse model is constructed
The core of the construction method of aggressive lymphomas mouse model of the invention is, traditional spontaneous B cell is drenched
It is small to obtain aggressive lymphomas for UTX gene knockout in the B cell of bar tumor mouse (such as E μ-Myc transgenic mice), screening
Mouse model.The aggressive lymphomas mouse model is the higher spontaneous B cell lymphoma mouse model of grade malignancy.
Specifically, can on the basis of traditional E μ-Myc transgenic mice, that is, spontaneous B cell lymphoma mouse, by with
The mouse of B cell specific deletion UTX gene, which mates, to be bred, and screening obtains aggressive lymphomas mouse model.
As shown in Figure 1A, the breeding of aggressive lymphomas mouse model has some requirements, wherein E μ-Myc transgenic loci
It needs to be kept by the mating pattern of semizygote E μ-Myc hero mouse and wild female mouse.
Specifically, as shown in Figure 1A, the construction method of aggressive lymphomas mouse model, comprising steps of (1) female UTX
Wild-type mice of the gene by gene targeting modification mates with male E μ-Myc transgenic mice, obtains UTX gene and passes through base
Because of the E μ-Myc transgenic mice for modification of practicing shooting；(2) wild-type mice and male of the female UTX gene by gene targeting modification
The mating of Cre transgenosis tool mouse, obtains UTX gene special knock-out mice in B cell；(3) hero obtained in step (1) is taken
Property UTX gene by gene targeting modification E μ-Myc transgenic mice and step (2) in acquisition female UTX gene it is thin in B
Special knock-out mice mating, obtains the E μ-Myc transgenic mice that UTX gene specifically knocks out in B cell in born of the same parents, screens, obtains
Aggressive lymphomas mouse model.Wherein, the male mouse and two UTX gene copies quilt that a UTX gene copy is knocked
The female mice of knockout all has the potentiality as aggressive lymphomas mouse model.
Wherein, building UTX gene by gene targeting modification wild-type mice when, can by DNA homologous recombination method,
Homologous sequence in section of DNA sequence containing LoxP sequence in targeting vector, with ES cells (embryonic stem cell) genome
It is replaced, to be inserted into LoxP sequence, Jin Erfan in No. 10 of UTX gene and No. 14 introne in mouse genome
It is bred as the wild-type mice by gene targeting modification.The building category of wild-type mice of the UTX gene by gene targeting modification
In the prior art, Liting Zheng, Longyong Xu.Qing Xu, Lu Yu, Danfeng Zhao, Pu Chen can refer to,
Wei Wang,Yiqin Wang,Gang Han,Charlie Degui Chen.Utx loss causes myeloid
transformation.Leukemia.2018Feb 20。
As shown in Figure 1B, another construction method of aggressive lymphomas mouse model, comprising steps of (1) female UTX base
Because the wild-type mice by gene targeting modification mates with male Cre transgenosis tool mouse, UTX gene is obtained in B cell
In special knock-out mice；(2) male E μ-Myc transgenic mice and step (1) middle female UTX gene obtained are special in B cell
Different knock-out mice mating, obtains the E μ-Myc transgenic mice that UTX gene specifically knocks out in B cell, and screening obtains invasion
Lymphoma mouse model.Wherein, what the male mouse and only one UTX gene copy that a UTX gene copy is knocked were knocked
Female mice all has the potentiality as aggressive lymphomas mouse model.
As shown in Figure 1 C, wild-type mice and male E μ-Myc transgenosis of the female UTX gene by gene targeting modification
Mouse mating obtains the E μ-Myc transgenic mice that UTX gene passes through gene targeting modification, as experiment contrast.
Two, the tumorigenic feature and life span of aggressive lymphomas mouse model
To acquired results in breeding scheme shown in breeding scheme shown in breeding scheme shown in above-mentioned Figure 1A, Figure 1B, Fig. 1 C point
It is not counted, as a result as shown in table 1-1, table 1-2 and table 1-3:
The mouse birth ratio of table 1-1 breeding scheme-Figure 1A
The mouse birth ratio of table 1-2 breeding scheme-Figure 1B
The mouse birth ratio of table 1-3 breeding scheme-Fig. 1 C
As a result, it is found that breeding scheme shown in above-mentioned Figure 1A, breeding scheme shown in Figure 1B, breeding scheme shown in Fig. 1 C, gained
Mouse carries out genotype identification and statistics, obtains the birth quantity of each mouse genotypes, the birth ratio of these mouse meets
Mendel's law illustrates these gene modifications and knocks out to influence very little for mouse early development.
In addition, we are to the target gene type mouse of breeding scheme acquisition and crt gene type in Figure 1A, Figure 1B and Fig. 1 C
Mouse is normally raised, and mouse invasion situation is observed, by carrying out touch detection to mouse axillary lymph knot weekly, to mouse
Spontaneous lymphoma there is a situation where being monitored, and birth and date of death are recorded, according to the existence of each mouse genotypes
Time draws survivorship curve figure, has obtained the result of Fig. 2.Target gene type mouse is as shown in blue line in Fig. 2 and green line, blue line point
E μ-Myc hero the mouse that a UTX gene copy knocks out and the E μ-Myc female mice that two UTX gene copies knock out, green line generation are not represented
E μ-Myc the female mice that one UTX gene copy of table knocks out, crt gene type mouse is as shown in red line in Fig. 2 and black line, red line generation
E μ-Myc hero the mouse of table UTX gene modification and female mice, black line represent male mouse and the female mice of UTX gene modification.
E μ-Myc the mouse that target gene type mouse, i.e. UTX gene are knocked in B cell, writes a Chinese character in simplified form into E μ-Myc；UTX
KO, shares 3 kinds of genotype classes, and the disease that these three mouse genotypes all have spontaneous generation malignant invasion lymthoma occurs
Potentiality.This 3 kinds of genotype details are as follows:
Genotype is expressed as E μ-Myc；UTXf/y；CD19-Cre- /+, write a Chinese character in simplified form into E μ-Myc；UTX-/y, i.e. a UTX base
Because of the E μ-Myc hero mouse that copy is knocked, it is shown in Table 1-1 and table 1-2, corresponding to the blue line in Fig. 2 hero mouse survivorship curve.
Genotype is expressed as E μ-Myc；UTXf/f；CD19-Cre- /+, write a Chinese character in simplified form into E μ-Myc；UTX-/-, i.e. two UTX bases
Because copying the E μ-Myc female mice being knocked, it is shown in Table 1-1, corresponding to the blue line in Fig. 2 female mice survivorship curve.
Genotype is expressed as E μ-Myc；UTXf/+；CD19-Cre- /+, write a Chinese character in simplified form into E μ-Myc；UTX- /+, i.e. a UTX base
Because of the E μ-Myc female mice that copy is knocked, it is shown in Table 1-2, corresponding to the green line in Fig. 2 female mice survivorship curve.
E μ-Myc the mouse and UTX gene that crt gene type mouse, i.e. UTX gene are modified are knocked in B cell
Mouse, write a Chinese character in simplified form into E μ-Myc respectively；UTXf/y and UTX-/y (male mouse), E μ-Myc；UTXf/f and UTX-/- (female mice), it is corresponding
4 kinds of genotype classes, details are as follows:
Genotype is expressed as E μ-Myc；E μ-Myc hero the mouse that UTXf/y, i.e. a UTX gene copy are modified, is shown in Table
1-3, corresponding to the red line in Fig. 2 hero mouse survivorship curve.
Genotype is expressed as E μ-Myc；E μ-Myc the female mice that UTXf/f, i.e. two UTX gene copies are modified, is shown in
Table 1-3, corresponding to the red line in Fig. 2 female mice survivorship curve.
Genotype is expressed as UTXf/y；CD19-Cre- /+, UTX-/y is write a Chinese character in simplified form into, i.e., a UTX gene copy is in B cell
In the male mouse that is knocked, 1-1 and table 1-2 are shown in Table, corresponding to the black line in Fig. 2 hero mouse survivorship curve.
Genotype is expressed as UTXf/f；CD19-Cre- /+, write a Chinese character in simplified form into UTX-/-, i.e., two UTX gene copies are in B cell
In the female mice that is knocked, 1-1 is shown in Table, corresponding to the black line in Fig. 2 female mice survivorship curve.
Supplementary explanation, the E μ-Myc mouse that UTX gene is modified are similar with traditional E μ-Myc mouse, it may occur that spontaneous
Lymthoma, but the grade malignancy of the spontaneous lymphoma of these mouse generation is very low, and lymthoma is seldom invaded to the outer organ of lymph node,
And obvious angiogenesis is had no around lymthoma, and the mouse survival and eugonic that UTX gene is knocked in B cell,
Have no the genius morbi in terms of lymthoma.
As shown in Fig. 2, compared with aggressive lymphomas mouse model control mice obtained by Fig. 1 C breeding scheme, in lymthoma
There is very big raising on proportion and lymthoma time of origin, in female mice, shows that lymthoma is UTX agent
Measure dependence-producing property.The tumorigenic feature and life span of male and each genotype of female mice are respectively illustrated in Fig. 2.
Three, feature occurs in situ for the lymthoma of each mouse genotypes in aggressive lymphomas mouse model
When obvious enlargement occurs for the axillary lymph tumor for touching mouse and morbid state occurs in mouse, mouse is implemented peaceful and comfortable
Extremely, and to mouse corpse it is dissected and is taken pictures, and the size of the spleen of mouse is measured and photographed to record, respectively obtained
The result of A, B, C in Fig. 3.Further, the spleen of mouse is fixed and is embedded, make paraffin section and HE dyeing, it is right
Centrum germinativum's structure in spleen carries out histologic analysis, has obtained the result of D in Fig. 3.Detailed process is as follows, HE dyeing group
The method of analysis includes the following steps: that cervical dislocation puts to death mouse, takes mouse spleen and lymthoma, is immersed in Ka Nuoshi fixer
In 4 DEG C of fixations overnight, be dehydrated using n-butanol, 12 hours every time, totally 4 times, waxdip 24 hours every time, totally 3 times, paraffin embedding
Afterwards, the ultra-thin paraffin section of 5um is made, is dyed using haematoxylin Yihong (HE) staining kit (the green skies C0105#), contaminates
Color step carries out to specifications, dewaxes through dimethylbenzene gradient, graded ethanol rehydration, and hematoxylin room temperature dyes ten minutes, 10mM
NaOH solution, which is impregnated several seconds, becomes blue to histotomy, and after impregnating 95% ethyl alcohol, eosin stains 1-2 minutes, 95% ethyl alcohol was clear
Eosin stain, dehydrated alcohol dehydration, dimethylbenzene is transparent, neutral gum mounting.Coloration result is just being set glimmering using OLYMPUS BX51
Light microscope is observed and is taken pictures.
To the mouse that lymthoma infringement head occurs, head epidermis is eliminated, takes pictures, is schemed to skull side
The result of E, F in 3.Intrathoracic lymphoma tissue and lung tissue are taken out, are clapped by the mouse for invading lymthoma thoracic cavity
According to having obtained the result of G, H in Fig. 3.Statistics and analysis is carried out to the quantity that the mouse that lymthoma invades head and thoracic cavity occurs,
The result of I in Fig. 3 is obtained.To photographing to record for a large amount of angiogenesis has occurred around the lymthoma of mouse, obtain
The result of J, K in Fig. 3.
As shown in figure 3, the performance of human malignant's lymthoma is extremely complex, including it is easy to recur and central nervous system occurs
Infringement and thymus gland infringement etc., the series of features which shows and human malignant's lymthoma
Feature is very similar.Figure A shows that feature occurs in situ for the lymthoma of each mouse genotypes, it is found that E μ-Myc；UTX KO
The blood vessel of the lymthoma of mouse becomes apparent from, and figure B and figure C show that the Spleen Size of each mouse genotypes, figure D show each gene
The HE coloration result of the spleen tissue of type mouse, E μ-Myc；The centrum germinativum of the spleen of UTX KO mouse destroys more serious, figure E
E μ-Myc is shown with figure F；Brain infringement has occurred in the lymthoma of UTX KO mouse, and figure G and figure H show E μ-Myc；UTX KO is small
Thymus gland infringement has occurred in the lymthoma of mouse, and figure I shows that brain and thymus gland, which occurs, invades the proportion in each genotype, schemes J
E μ-Myc is shown with figure K；Apparent angiogenesis phenomenon has occurred in the lymthoma of UTX KO mouse.
Four, feature occurs for the lymthoma of aggressive lymphomas mouse model
Due to the especially complex multiplicity of the characterization of spontaneous lymphoma model, we further present a part of aggressive lymph
Feature occurs for the lymthoma of tumor mouse model, and wherein Fig. 4 A is the supplement of Fig. 3 E, F, and Fig. 4 B is the supplement of Fig. 3 G, H, and Fig. 4 C, D are
The supplement of Fig. 3 J, K, and Fig. 4 E is that the lymthoma of some animals invades the picture in abdominal cavity.
As shown in figure 4, these supplement pictures more fully demonstrate the lymthoma of the aggressive lymphomas mouse model
Feature occurs.Figure A and figure B show E μ-Myc；The example that brain and thymus gland are invaded occurs for the lymthoma of UTX KO mouse, and figure C is aobvious
E μ-Myc is shown；Apparent angiogenesis phenomenon has occurred in the lymthoma of UTX KO mouse, and figure D shows E μ-Myc；UTX KO
The in-situ image of apparent angiogenesis phenomenon has occurred in the lymthoma of mouse, and figure E shows E μ-Myc；The leaching of UTX KO mouse
Abdominal cavity infringement has occurred in bar tumor.
Five, the origin of cell analysis of aggressive lymphomas mouse model:
We separate the lymthoma that each mouse genotypes occur, and are dyed by fluidic cell and carry out cell surface
Developed by molecule analysis, which for being initiated by B cell differentiation and development to lymthoma are analyzed in stage, have obtained the knot in Fig. 5
Fruit.Wherein, flow cytometry includes the following steps: the spleen, marrow and lymthoma that take corresponding mouse respectively, buffers in streaming
In liquid (1%BSA PBS), tissue is squeezed using sheet glass hair side, cell suspension is made, obtained using strainer filtering cell slender
Born of the same parents' suspension after centrifugation, is resuspended cell using erythrocyte cracked liquid (ACK), splits in 37 DEG C of water-baths red 5 minutes, after centrifugation, make
Cell is resuspended with streaming buffer, using Flow cytometry, takes 1x106 cell to manage to new FACS, it is slow that 0.5ml streaming is added
Cell is resuspended in fliud flushing, and 0.2ul is added after centrifugation, while in every 30ul streaming buffer and corresponds to antibody, antibody dyeing liquor is made, often
A sample is resuspended using 30ul antibody dyeing liquor, is put into 4 DEG C of refrigerators and is protected from light dyeing 40 minutes to 1 hour.It is buffered using 3ml streaming
Liquid dilutes antibody dyeing liquor, after centrifugation, sample is resuspended using 200ul buffer, is detected using flow cytometer.
The origin of cell of human lymphoma and the prognosis of lymthoma and closely related, the most of mankind's malignant lymphomas for the treatment of
Originating from mature B cell.By fluidic cell surface molecular labeled analysis, as shown in figure 5, aggressive lymphomas of the invention is small
The cell of mouse model is originating primarily from the later period of bone-marrow-derived lymphocyte development, similar to the origin of cell of human lymphoma.Figure A is shown
The origin of cell distribution proportion of the lymthoma of each mouse genotypes comes, and figure B is that the origin of cell molecular labeling of bone-marrow-derived lymphocyte shows
It is intended to.
The above, only presently preferred embodiments of the present invention, not to the present invention in any form with substantial limitation,
It should be pointed out that under the premise of not departing from the method for the present invention, can also be made for those skilled in the art
Several improvement and supplement, these are improved and supplement also should be regarded as protection scope of the present invention.All those skilled in the art,
Without departing from the spirit and scope of the present invention, when made using disclosed above technology contents it is a little more
Dynamic, modification and the equivalent variations developed, are equivalent embodiment of the invention；Meanwhile all substantial technologicals pair according to the present invention
The variation, modification and evolution of any equivalent variations made by above-described embodiment, still fall within the range of technical solution of the present invention
It is interior.
Claims (10)
1. a kind of construction method of aggressive lymphomas mouse model includes the following steps: spontaneous B cell lymphoma mouse
UTX gene knockout, screening obtain aggressive lymphomas mouse model.
2. the construction method of aggressive lymphomas mouse model according to claim 1, which is characterized in that spontaneous B is thin
UTX gene knockout in born of the same parents' Lymphoma Mice B cell.
3. the construction method of aggressive lymphomas mouse model according to claim 1, which is characterized in that use gene work
The method of journey knocks out UTX gene.
4. the construction method of aggressive lymphomas mouse model according to claim 1, which is characterized in that the spontaneous B
Cell lymphoma mouse is E μ-Myc transgenic mice.
5. the construction method of aggressive lymphomas mouse model according to claim 1, which is characterized in that spontaneous B is thin
Born of the same parents' Lymphoma Mice mates breeding with the mouse of B cell specific deletion UTX gene, and screening obtains aggressive lymphomas from offspring
Mouse model.
6. the construction method of aggressive lymphomas mouse model according to claim 1, which is characterized in that including walking as follows
It is rapid:
(1) wild-type mice of the female UTX gene by gene targeting modification mates with male E μ-Myc transgenic mice, obtains
UTX gene passes through the E μ-Myc transgenic mice of gene targeting modification；(2) female UTX gene passes through the open country of gene targeting modification
Raw type mouse mates with male Cre transgenosis tool mouse, obtains UTX gene special knock-out mice in B cell；(3) step is taken
Suddenly acquisition is female in E μ-Myc transgenic mice and step (2) of the male UTX gene obtained in (1) by gene targeting modification
Property the special knock-out mice mating in B cell of UTX gene, obtain the E μ-Myc transgenosis that UTX gene specifically knocks out in B cell
Mouse, screening obtain aggressive lymphomas mouse model.
7. the construction method of aggressive lymphomas mouse model according to claim 1, which is characterized in that including walking as follows
It is rapid:
(1) wild-type mice of the female UTX gene by gene targeting modification mates with male Cre transgenosis tool mouse, obtains
Obtain UTX gene special knock-out mice in B cell；(2) male E μ-Myc transgenic mice and step (1) middle female obtained
The special knock-out mice mating in B cell of UTX gene, it is small to obtain the E μ-Myc transgenosis that UTX gene specifically knocks out in B cell
Mouse, screening obtain aggressive lymphomas mouse model.
8. the aggressive lymphomas mouse model that the construction method as described in any one of claim 1~7 prepares.
9. construction method or aggressive lymphomas mouse mould as claimed in claim 8 as described in any one of claim 1~7
Type is used to screen the purposes of aggressive lymphomas therapeutic agent.
10. a kind of method for screening aggressive lymphomas therapeutic agent, comprising steps of being applied to candidate substances such as claim
The aggressive lymphomas mouse model that 1~7 any one construction method obtains can incite somebody to action if aggressive lymphomas weakens
The drug candidate is as candidate aggressive lymphomas therapeutic agent.
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Cited By (2)
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|---|---|---|---|---|
| CN115281153A (en) * | 2022-08-30 | 2022-11-04 | 上海米地生物医药有限公司 | Construction and application of primary central nervous system lymphoma mouse model |
| CN115399292A (en) * | 2022-04-01 | 2022-11-29 | 上海米地生物医药有限公司 | Construction and application of primary extralymph node lymphoma mouse model |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060117399A1 (en) * | 2004-11-30 | 2006-06-01 | Medical University | Animal model for human lymphomas |
| CN106244552A (en) * | 2015-06-05 | 2016-12-21 | 上海交通大学医学院附属瑞金医院 | The method for building up of a kind of diffusivity large B cell type lymphoma animal models and application thereof |
-
2018
- 2018-05-04 CN CN201810420761.0A patent/CN110438158B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060117399A1 (en) * | 2004-11-30 | 2006-06-01 | Medical University | Animal model for human lymphomas |
| CN106244552A (en) * | 2015-06-05 | 2016-12-21 | 上海交通大学医学院附属瑞金医院 | The method for building up of a kind of diffusivity large B cell type lymphoma animal models and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| GIJS VAN HAAFTEN ET AL.: ""Somatic mutations of the histone H3K27 demethylase, UTX, in human cancer"", 《NAT GENET.》 * |
| JONI VAN DER MEULEN ET AL.: ""The H3K27me3 demethylase UTX is a gender-specific tumor suppressor in T-cell acute lymphoblastic leukemia"", 《BLOOD》 * |
| LING TIAN ET AL.: ""The Complex Role of KDM6A in B-Cell Development and Function"", 《BLOOD》 * |
| XIAOXI LI ET AL.: ""UTX is an escape from X-inactivation tumorsuppressor in B cell lymphoma suppressor in B cell lymphoma"", 《NATURE COMMUNICATIONS》 * |
| 杨贇 等: ""组蛋白H3K27去甲基化酶UTX在肿瘤研究中的作用"", 《中国老年学杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115399292A (en) * | 2022-04-01 | 2022-11-29 | 上海米地生物医药有限公司 | Construction and application of primary extralymph node lymphoma mouse model |
| CN115399292B (en) * | 2022-04-01 | 2024-04-02 | 上海米地生物医药有限公司 | Construction and application of primary extra-lymph node lymphoma mouse model |
| CN115281153A (en) * | 2022-08-30 | 2022-11-04 | 上海米地生物医药有限公司 | Construction and application of primary central nervous system lymphoma mouse model |
| CN115281153B (en) * | 2022-08-30 | 2024-04-05 | 上海米地生物医药有限公司 | Construction and application of primary central nervous system lymphoma mouse model |
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