CN106591446A - Application of cellular pathway regulation molecules as drug target and to EBOV (Ebola virus) infection diagnosis - Google Patents
Application of cellular pathway regulation molecules as drug target and to EBOV (Ebola virus) infection diagnosis Download PDFInfo
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Abstract
The invention discloses application of cellular pathway regulation molecules as a drug target and to EBOV (Ebola virus) infection diagnosis. The cellular pathway regulation molecules refer to TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3, MSR1, MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2, MTOR, CASP8 and LAM2. The 22 molecules can be used as key target molecules for designing anti-EBOV infection drugs and guiding strategic selection of clinical intervention means, and can also be used as biomolecule identification for clinical diagnosis of EBOV infected persons.
Description
Technical field
The present invention relates in biological technical field, cell pathway regulatory molecule is as drug target and diagnosis EBOV infection
In application.
Background technology
Ebola disease viral disease is by the Ebola virus (Ebola virus, EBOV) of Filoviridae (filoviridae)
A kind of caused acute hemorrhage sexually transmitted disease.Case fatality rate is very high, up to 50%～90%.
Ebola virus genome is sub-thread strand RNA, and about long 19kb can encoding nuclear proteins (NP) and envelope protein
7 structural protein such as (VP35, VP40, VP30, VP24), glycoprotein (GP) and RNA polymerase, wherein GP gene pairss EBOV are replicated
There are the coding and functional transcription of uniqueness.Virus is replicated in the kytoplasm of infection cell, assembling, is discharged in bud life mode.Ebola
It is viral mainly to include Zaire's type (EBOV-Z), the Sudan's type (EBOV-S), Cote d'lvoire's type (EBOV-C) and Reston type
(EBOV-R) etc., the characteristic of different subtype is different, and wherein Zaire's type virulence is most strong, and the Sudan's type takes second place, and both are to the mankind and non-
The fatality rate of people primatess is very high.Reston type and Cote d'lvoire's type are relatively low to the virulence of people, show as subclinical infection, but right
Non-human primates have mortality.
EBOV is a kind of Zoonosis virus, and its route of transmission is still not clear, generally by with mankind's close contact to infecting
The blood of animal, secretions, organ or other body fluid cause infection, and body fluid of directly contact the infected etc. can cause interpersonal
Propagate.Various non-human primate and the mankind are generally susceptible.Therefore, family member of HIV-infected people, medical worker, reviewer, angstrom
The rich staff for drawing viral prevalence scene, infection animal contactee etc. are high-risk group.
EBOV is a kind of virus of pantropic, can invade each system organ, is attached most importance to liver, spleen infringement especially.The generation of primary disease
It is relevant with the immunne response level of body.Especially phagocyte is thin by the target of virus attack first to mononuclear phagocyte system
Born of the same parents, subsequent fibroblast and endotheliocyte it is infected, vascular permeability increases, and fibrin is calm.After infection 2 days it is viral
Detect first in lung, detect in the tissue such as liver, spleen after 4 days, 6 Tian Hou body tissues can detect.
Many researchs with regard to pathogenicity at present are all carried out in animal model and experiment in vitro, and with regard to people
The correlational study of precursor virus infection is few.Each stage of virus infection body, from cell entry cell, partial copy, propagation (disease
Toxemia), in the target organ breeding, Rehabilitation (but still band poison) or dead, there is the mutual restriction of two kinds of factors：Virus
Infection and host body response and defence.Research pathogenesis and Immune escaping mechanism of the EBOV to the mankind, and then according to these
Committed step, determines modulation site, researches and develops medicine, for the Synthetical prevention of ebola hemorrhagic fever has important meaning
Justice.Meanwhile, the response feature of human infection EBOV is studied, the molecular marker of infection and prognosis characterizations is found, it is fast for setting up
Fast diagnostic method, set up timely and effective intervention stratege theoretical foundation can be provided.
The content of the invention
The technical problem to be solved is how to diagnose EBOV infection and how to design treatment and/or prevent EBOV
The drug target of infection.
For solve above-mentioned technical problem, present invention firstly provides following R1)-R4) in arbitrary application：
R1) G1 is preparing treatment and/or is preventing the application in EBOV infection products as target spot；
R2) G1 is being treated and/or is being prevented the application in EBOV infection as target spot；
R3) detect application of the system of the G1 expressions in diagnosis or auxiliary diagnosis EBOV infection product is prepared；
R4) detect application of the system of the G1 expressions in diagnosis or the infection of auxiliary diagnosis EBOV；
The G1 be TRAF2 genes, TP53 genes, CASP9 genes, RIPK3 genes, CASP7 genes, CIAPIN1 genes,
Tnf gene, BAX genes, CASP6 genes, TYRO3 genes, MSR1 genes, MARCO genes, HAVCR1 genes, IRS1 genes,
LRSAM1 genes, TPCN2 genes, TPCN1 genes, ULK1 genes, AKT2 genes, MTOR genes, CASP8 genes and/or LAMP2
Gene.
In above-mentioned application, the system of the detection G1 expressions can be to detect the G1 using method of the prior art
Reagent and/or instrument needed for expression, reagent as described in being detected using high-flux sequence needed for G1 expressions and/or
Instrument, or using the reagent and/or instrument needed for G1 expressions described in quantitative PCR detection, or hybridize skill using northern
Art detects the reagent and/or instrument needed for the G1 expressions, or using needed for G1 expressions described in genechip detection
Reagent and/or instrument.
In above-mentioned application, the high-flux sequence can be carried out using Illumina microarray datasets.
In above-mentioned application, the system of the detection G1 expressions may also include data handling system, the data processing
System is used for analyzing the result that method of the prior art is directly obtained, and determines the expression of the G1.The data processing
System can be software and/or module.
For solve above-mentioned technical problem, present invention also offers following M1)-M4) in arbitrary application：
M1) P1 is preparing treatment and/or is preventing the application in EBOV infection products as target spot；
M2) P1 is being treated and/or is being prevented the application in EBOV infection as target spot；
M3) detect application of the system of the P1 contents in diagnosis or auxiliary diagnosis EBOV infection product is prepared；
M4) detect application of the system of the P1 contents in diagnosis or the infection of auxiliary diagnosis EBOV；
The P1 be TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3,
MSR1, MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2, MTOR, CASP8 and/or LAMP2 albumen
Matter.
In above-mentioned application, the system of the detection P1 contents can be to detect the P1 contents using method of the prior art
Required reagent and/or instrument, the reagent and/or instrument needed for using mass spectrum or its P1 content as described in correlation technique detection,
Or using the reagent and/or instrument needed for P1 contents described in immuning hybridization technology for detection, or the P1 is detected using elisa technique
Reagent and/or instrument needed for content, or using the reagent and/or instrument needed for P1 contents described in protein chip or detection paper
Device.
To solve above-mentioned technical problem, present invention also offers following N1) or application N2)：
N1) using above-mentioned G1 as system used in the diagnosis of EBOV infection marks or auxiliary diagnosis EBOV infection method
In following a1) or a2) in application：
A1 diagnosis or auxiliary diagnosis EBOV infection product) are prepared；
A2) diagnosis or the infection of auxiliary diagnosis EBOV；
N2) using above-mentioned P1 as system used in the diagnosis of EBOV infection marks or auxiliary diagnosis EBOV infection method
In above-mentioned a1) or a2) in application.
It is described using above-mentioned G1 as EBOV infect mark diagnosis or auxiliary diagnosis EBOV infection method in used be
System can be the system of detection G1 expressions mentioned above.
To solve above-mentioned technical problem, present invention also offers following W1) or product W2)：
W1 EBOV infection products) are treated and/or prevent, the product is with above-mentioned G1 as target spot；
W2 EBOV infection products) are treated and/or prevent, the product is with above-mentioned P1 as target spot.
To solve above-mentioned technical problem, present invention also offers following X1) or product X2)：
X1) diagnosis or auxiliary diagnosis EBOV infection product, are the system for detecting above-mentioned G1 expressions；
X2) diagnosis or auxiliary diagnosis EBOV infection product, are the system for detecting above-mentioned P1 contents.
To solve above-mentioned technical problem, present invention also offers the EBOV infection relevant molecular characteristics spectrums of non-diagnostic purpose
Construction method.
The construction method of the EBOV infection relevant molecular characteristics spectrums of non-diagnostic purpose provided by the present invention includes：It is right respectively
RNA or protein in EBOV infected groups and non-EBOV infected groups blood carries out quantitative analyses, obtains EBOV according to analysis result
Virus load relevant molecular characteristics are composed；The EBOV virus loads relevant molecular characteristics spectrum is above-mentioned G1 or above-mentioned P1.
In said method, the RNA or protein in EBOV infected groups and non-EBOV infected groups blood is carried out quantitatively
Analysis specifically may also include：RNA or protein in purification EBOV infected groups and non-EBOV infected groups blood, using Illumina
Microarray dataset detects the content of RNA or protein.
RNA or protein in the purification EBOV infected groups and non-EBOV infected groups blood is using with Oligo
(dT) magnetic bead is carried out.
The content of the utilization Illumina microarray datasets detection RNA or protein may include：By RNA fragments after purification
Change, and as template reverse transcription synthetic double chain cDNA：End is carried out to double-strand cDNA and repairs polishing so as to 5 ' end phosphoric acid
Change, 3 ' ends add " A "；Double-strand cDNA joint with 3 ' dTMP ends is connected in sequence measuring joints, is expanded by PCR, using AMPure
XP enrichment with magnetic bead purification joint products, obtain library；The library for building is carried out double ends to survey with Illumina microarray datasets
Sequence, detects the content of RNA or protein in sample to be tested.
Above, the G1 expressions can be the expression of G1 described in blood.The P1 contents can be in blood
The content of the P1.
Above, the treatment and/or prevention EBOV infection product can be medicine or vaccine.The diagnosis or auxiliary diagnosis
EBOV infection product can be medicine and/or health equipment, such as reagent, reagent paper, material or instrument.
In actual applications, can by compare TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX,
CASP6, TYRO3, MSR1, MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2 and MTOR and
Expressions of the CASP8 and LAMP2 in object to be measured with non-EBOV the infected's blood is judging whether object to be measured is EBOV
The infected.Specific criterion is as follows：As the TRAF2 of object to be measured, TP53, CASP9, RIPK3, CASP7, CIAPIN1,
TNF, BAX, CASP6, TYRO3, MSR1, MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2 and MTOR
In the expression of at least one higher than 2 times or CASP8 and LAMP2 of the corresponding molecule of non-EBOV the infected at least one
Expression less than the corresponding molecule of non-EBOV the infected 1/2, the object to be measured be candidate be EBOV the infected；As described
The TRAF2 of object to be measured, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3, MSR1,
The expression of MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2 and MTOR is not higher than non-EBOV
2 times of the corresponding molecule of the infected and the expression of CASP8 and LAMP2 is not less than the 1/2 of the corresponding molecule of non-EBOV the infected,
The object to be measured is or candidate is non-EBOV the infected.
The present invention is by the non-EBOV the infected of comparison and the difference expression gene of EBOV the infected, and is divided by dependency
Analysis, finds the differential expression molecule (r related to EBOV virus loads in EBOV the infected's blood2>0.64,p<0.01)——
TRAF2、TP53、CASP9、RIPK3、CASP7、CIAPIN1、TNF、BAX、CASP6、TYRO3、MSR1、MARCO、HAVCR1、
IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2 and MTOR and CASP8 and LAMP2, this 22 molecules are infected in EBOV
The high expression of blood samples of patients cell can the efficient phagocytic virus granule of inducing host cell, but but cannot quickly remove virus,
So that virus is successfully, reproduced in the cell, and the virus for replicating is spread in vivo by necrosis and apoptosis.These result tables
The key molecule of bright these host cells participates in and decides the destiny of EBOV infected patient body cells, participates in EBOV in body
In course of infection, can be used as key target molecule, the design and guidance for anti-Ebola virus infection medicine are clinical dry
The policy selection of pre- means, is also used as biomolecule mark, for the clinical diagnosises of EBOV the infected.
Specific embodiment
The present invention is further described in detail with reference to specific embodiment, the embodiment for being given is only for explaining
The bright present invention, rather than in order to limit the scope of the present invention.
Experimental technique in following embodiments, if no special instructions, is conventional method.
In following embodiments, material used, reagent etc., if no special instructions, commercially obtain.
Embodiment 1, the host's key molecule for participating in EBOV infected patient cell regulate and controls
Total serum IgE is extracted from EBOV the infected with non-EBOV the infected's whole blood, it is pure with the absorption of the magnetic bead with Oligo (dT)
Change the mRNA in total serum IgE, in a heated condition by mRNA fragmentations, and as template reverse transcription synthetic double chain cDNA.To double
Chain cDNA carries out end and repairs polishing so as to 5 ' end phosphorylations, and 3 ' ends add " A ".Double-strand cDNA joint with 3 ' dTMP ends is connected
To in sequence measuring joints, expanded by PCR, using AMPure XP enrichment with magnetic bead purification joint products, by PCR reaction identification texts
Storehouse.The library for building is carried out into double end sequencings with Illumina microarray datasets.Filter what sequencing was produced using perl script
Joint sequence, two ends low quality sequence (Q in initial data<=20 base number accounts for more than the 50% of whole piece reads sequence
Row), Sequences of Low Complexity.
Valid data are compared into people with reference on genome using Tophat softwares, Cutfflinks software combinations compare knot
Really, differential expression of the EBOV the infected (n=22) compared with non-EBOV the infected (n=10) is drawn by Cutffdiff softwares
Gene, further by correlation analysiss, finds to infect the biomolecule for determining that host cell destiny is relevant with EBOV.As a result show
Show, 22 host's key molecules regulate and control closely related (p with the caused cell pathway of EBOV infection<0.0001, differential expression multiple
>2；Except TYRO3, the p value of TYRO3 is for 0.0024), this 22 molecules are respectively：Apoptosis correlation molecule TNF, TP53,
CASP9, CASP7, BAX, CASP6 and CASP8；Necrocytosiss correlation molecule TNF, TRAF2, CIAPIN1 and RIPK3；Cell is gulped down
Bite associated receptor TYRO3, MSR1, MARCO, HAVCR1, TPCN2 and TPCN1；Cell autophagy correlation molecule IRS1, AKT2,
TP53, MTOR, ULK1, LRSAM1 and LAMP2；Wherein, except CASP8 and LAMP2 are presented significant table in EBOV the infected
Outer up to lowering, other molecules occur significant up-regulated in EBOV the infected, and concrete outcome as shown in table 1, shows this
22 molecules can be identified as biomolecule, for the clinical diagnosises of EBOV the infected.
Table 1 determines the key biological molecule of EBOV virus host cells infected destiny
Note：22 molecules are listed in table 1 in EBOV the infected relative to differential expression multiple in non-EBOV the infected
(median FKPM values) and Variant statistical analysis result (P values).Wherein, TRAF2 is TNF receptor associated
The abbreviation of factor 2, abbreviations of the TP53 for tumor protein p53；Abbreviations of the CASP9 for caspase 9；RIPK3 is
The abbreviation of receptor-interacting serine-threonine kinase 3；Abbreviations of the CASP7 for caspase 7；
Abbreviations of the CIAPIN1 for cytokine induced apoptosis inhibitor 1；TNF is tumor necrosis
The abbreviation of factor；Abbreviations of the BAX for BCL2 associated X；Abbreviations of the CASP6 for caspase 6；TYRO3 is TYRO3
The abbreviation of protein tyrosine kinase 3；Abbreviations of the MSR1 for macrophage scavenger receptor 1；
Abbreviations of the MARCO for macrophage receptor with collagenous structure；HAVCR1 is hepatitis
The abbreviation of A virus cellular receptor 1；Abbreviations of the IRS1 for insulin receptor substrate 1；
Abbreviations of the LRSAM1 for leucine rich repeat and sterile alpha motif containing 1；TPCN2
For the abbreviation of two pore segment channel 2；Abbreviations of the TPCN1 for two pore segment channel 1；
Abbreviations of the ULK1 for unc-51 like autophagy activating kinase 1；AKT2 is AKT serine/
The abbreviation of threonine kinase 2；Abbreviations of the MTOR for mechanistic target of rapamycin；CASP8 is
The abbreviation of caspase 8；Abbreviations of the LAMP2 for lysosomal-associated membrane protein 2.
In actual applications, can by compare TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX,
CASP6, TYRO3, MSR1, MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2 and MTOR and
The expression of CASP8, LAMP2 in object to be measured with non-EBOV the infected's blood is come whether judge object to be measured be EBOV senses
Dye person.Specifically, the criterion for being obtained according to the above results is as follows：As the TRAF2 of object to be measured, TP53, CASP9,
RIPK3、CASP7、CIAPIN1、TNF、BAX、CASP6、TYRO3、MSR1、MARCO、HAVCR1、IRS1、LRSAM1、TPCN2、
In TPCN1, ULK1, AKT2 and MTOR the expression of at least one higher than 2 times of the corresponding molecule of non-EBOV the infected or
The expression of at least one less than the corresponding molecule of non-EBOV the infected 1/2 in CASP8 and LAMP2, the object to be measured are doubted
Like EBOV the infected；As the TRAF2 of object to be measured, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6,
The expression of TYRO3, MSR1, MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2 and MTOR is not
Higher than 2 times of the corresponding molecule of non-EBOV the infected and the expression of CASP8 and LAMP2 to be not less than non-EBOV the infected corresponding
The 1/2 of molecule, the object to be measured are non-EBOV the infected.
TRAF2、TP53、CASP9、RIPK3、CASP7、CIAPIN1、TNF、BAX、CASP6、TYRO3、MSR1、MARCO、
HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2 and MTOR and CASP8 and LAMP2 this 22 molecules exist
The high expression of EBOV infected patient blood cells can the efficient phagocytic virus granule of inducing host cell, but but cannot be quickly clear
Except virus so that virus is successfully, reproduced in the cell, and the virus for replicating is spread in vivo by necrosis and apoptosis.These
As a result show that the key molecule of these host cells participates in and decide the destiny of EBOV infected patient body cells, participate in EBOV
Course of infection in body, can be as key target molecule, for the design and guidance of anti-Ebola virus infection medicine
The policy selection of clinical intervention means.
The present invention also have chosen the blood sample of 6 non-EBOV the infecteds and 20 EBOV the infecteds in addition, using fixed
Amount RT-PCR technology have detected the table of RIPK3 genes and Casp6 genes in this 22 molecules in these object of study blood
Up to level.Wherein, the primer sequence of RIPK3 genes be 5 '-CGGGCGCAACATAGGAAGT-3 ' (sequence 1) and 5 '-
CTTCTAGGCGCAGCACGAAT-3 ' (sequence 2), the primer sequence of Casp6 genes is 5'-
GTCAACTGTTAGCCACGCAGAT-3'(sequences are 3) with 5'-AACCAGGCTGTGACACTTGTCT-3'(sequences 4).
As a result find, the expression of RIPK3 genes and Casp6 genes in EBOV the infected's blood is substantially less than non-
In EBOV the infected's blood, corresponding Cytokine Expression Level (table 2), shows, RIPK3 and Casp6 can be auxiliary as mark
Diagnosis EBOV the infected is helped, key target molecule is also used as, design for anti-Ebola virus infection medicine and is referred to
Lead the policy selection of clinical intervention means.
Table 2, cell death associated gene (RIPK3, CASP6) RT-PCR experimental results
Embodiment 2, the tiny RNA s (sRNAs) for participating in EBOV infection damage regulation and control
Total serum IgE is extracted from EBOV the infected with non-EBOV the infected's whole blood, using 15% denaturing polyacrylamide gel
Electrophoretic techniquess (PAGE) separate small fragment (15-30nt) RNA from total serum IgE, after purification to detached small fragment RNA connection 3 ' and
5 ' end connectors, then the RNA for being connected with joint is inverted with SuperScriptII reverse transcription (Invitrogen products)
Record, synthesizes cDNA.Then, performing PCR amplification, amplification program are entered：98 DEG C of denaturations 30s, 98 DEG C of degeneration 10s, 60 DEG C of annealing 30s,
Thus 72 DEG C of extension 15s, 14 circulations, 72 DEG C of extension 8min build library.Finally, product utilization Illumina sequencings are extended
Platform carries out high-flux sequence.Joint sequence, the two ends low quality being sequenced in the initial data for producing is filtered using perl script
Sequence (sQ<=5 base number accounts for the sequence of more than the 50% of whole piece reads sequence, the ratio of N more than 10%, polyA/T/
G/C sequences).
SRNA valid data after screened by length compare people with reference on genome, analyze
Distribution and expression of the sRNAs in reference gene group, the sequence in comparison is compared with miRBase sequences, obtains
MiRNA information, while removing rRNA, tRNA, snRNA, snoRNA as far as possible.EBOV the infected (n is drawn with reference to sample information
=differential expression miRNA 12) compared with non-EBOV the infected (n=3), further by correlation analysiss, has found and EBOV
The related miRNA of virus infection.As a result show, belong to 5 miRNA (hsa- in 1 miRNA family (hsa-let-7 families)
Let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-5p and hsa-let-7f-5p), they
(p is remarkably decreased relative to expression in non-EBOV infection Healthy Peoples in EBOV the infected<0.01, differential expression multiple>4), and
Their target gene significantly increases (p accordingly<0.0001, differential expression multiple>3), concrete outcome is as shown in table 3, table 4.
Hsa-let-7 families miRNAs can regulate and control immunne response correlation molecule IL6, IGDCC3, hepatic injury correlation molecule CHRD, cell
Dead correlation molecule DPP6, FASLG, cell cycle and development correlation molecule E2F5, E2F6, MAB21L3, MEIS3, tissue repair
The gene expression of correlation molecule LIN28 etc., these results show 5 miRNA (hsa-let-7a- in hsa-let-7 families
5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-5p and hsa-let-7f-5p) and its target gene
(MAB21L3, MEIS3, DPP6, IGDCC3, CHRD, FASLG, LIN28, E2F5, IL6 and E2F6) can be used as biomolecule mark
Know, for the clinical diagnosises of EBOV the infected, it is also possible to as biological target molecule, for the design of anti-EBOV infection medicines.
Wherein, MAB21L3, MEIS3, DPP6, IGDCC3, CHRD, FASLG, LIN28, E2F5, IL6 and E2F6 are
The target base of hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-5p and hsa-let-7f-5p
Cause.
In actual applications, can be by comparing 5 miRNA (hsa-let-7a-5p, hsa- in hsa-let-7 families
Let-7b-5p, hsa-let-7c-5p, hsa-let-7d-5p and hsa-let-7f-5p) and its target gene (MAB21L3,
MEIS3, DPP6, IGDCC3, CHRD, FASLG, LIN28, E2F5, IL6 and E2F6) in object to be measured and non-EBOV the infected's blood
Expression in liquid is judging whether object to be measured is EBOV the infected.Specifically, the judgement mark for being obtained according to the above results
It is accurate as follows：As object to be measured hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-5p and
In hsa-let-7f-5p the expression of at least one less than the corresponding molecule of non-EBOV the infected 1/4 or MAB21L3,
In MEIS3, DPP6, IGDCC3, CHRD, FASLG, LIN28, E2F5, IL6 and E2F6, the expression of at least one is higher than non-
3 times of the corresponding molecule of EBOV the infected, the doubtful EBOV the infected of the object to be measured；The such as hsa-let-7a-5p of object to be measured,
The expression of hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-5p and hsa-let-7f-5p is not less than non-
1/4 and MAB21L3, MEIS3, DPP6, IGDCC3, CHRD, FASLG, LIN28, E2F5, IL6 of the corresponding molecule of EBOV the infected
3 times of non-EBOV the infected corresponding molecule are not higher than with the expression of E2F6, the object to be measured is non-EBOV the infected.
Table 3, the miRNAs for participating in EBOV infection damage regulation and control
Note：Tables of 5 miRNA of hsa-let-7 families in EBOV the infected and non-EBOV the infecteds is listed in table 3
Up to level (median TPM values) and Variant statistical analysis result (P values).
Table 4, hsa-let-7 families miRNA target gene in EBOV the infected differential expression
Note：The target gene of hsa-let-7 families miRNA is listed in table 4 in EBOV the infected and non-EBOV the infecteds
Expression (median FKPM values) and Variant statistical analysis result (P values).Wherein, MAB21L3 is mab-21-like 3
Abbreviation；Abbreviations of the MEIS3 for Meis homeobox 3；Abbreviations of the DPP6 for dipeptidyl peptidase like 6；
IGDCC3 be immunoglobulin superfamily, DCC subclass, the abbreviation of member 3；CHRD is chordin
Abbreviation；Abbreviations of the FASLG for Fas ligand；Abbreviations of the LIN28 for lin-28 homolog；E2F5 is E2F
The abbreviation of transcription factor 5；Abbreviations of the IL6 for interleukin 6；E2F6 is E2F transcription
The abbreviation of factor 6.
<110>Inst. of Epidemiology and Microbiology, Academy of Military Medical Sciences, PL
<160> 4
<170> PatentIn version 3.5
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Claims (9)
1. following R1)-R4) in arbitrary application：
R1) G1 is preparing treatment and/or is preventing the application in EBOV infection products as target spot；
R2) G1 is being treated and/or is being prevented the application in EBOV infection as target spot；
R3) detect application of the system of the G1 expressions in diagnosis or auxiliary diagnosis EBOV infection product is prepared；
R4) detect application of the system of the G1 expressions in diagnosis or the infection of auxiliary diagnosis EBOV；
The G1 is TRAF2 genes, TP53 genes, CASP9 genes, RIPK3 genes, CASP7 genes, CIAPIN1 genes, TNF
Gene, BAX genes, CASP6 genes, TYRO3 genes, MSR1 genes, MARCO genes, HAVCR1 genes, IRS1 genes,
LRSAM1 genes, TPCN2 genes, TPCN1 genes, ULK1 genes, AKT2 genes, MTOR genes, CASP8 genes and/or LAMP2
Gene.
2. application according to claim 1, it is characterised in that：The system of the detection G1 expressions is the detection G1
Reagent and/or instrument needed for expression.
3. following M1)-M4) in arbitrary application：
M1) P1 is preparing treatment and/or is preventing the application in EBOV infection products as target spot；
M2) P1 is being treated and/or is being prevented the application in EBOV infection as target spot；
M3) detect application of the system of the P1 contents in diagnosis or auxiliary diagnosis EBOV infection product is prepared；
M4) detect application of the system of the P1 contents in diagnosis or the infection of auxiliary diagnosis EBOV；
The P1 be TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3, MSR1,
MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2, MTOR, CASP8 and/or LAMP2 protein.
4. application according to claim 3, it is characterised in that：Detect the system of the P1 contents to detect the P1 contents
Required reagent and/or instrument.
5. following N1)-N4) in arbitrary application：
N1) using G1 as system used in the diagnosis of EBOV infection marks or auxiliary diagnosis EBOV infection method following
A1 the application in) or a2)：
A1 diagnosis or auxiliary diagnosis EBOV infection product) are prepared；
A2) diagnosis or the infection of auxiliary diagnosis EBOV；
The G1 is TRAF2 genes, TP53 genes, CASP9 genes, RIPK3 genes, CASP7 genes, CIAPIN1 genes, TNF
Gene, BAX genes, CASP6 genes, TYRO3 genes, MSR1 genes, MARCO genes, HAVCR1 genes, IRS1 genes,
LRSAM1 genes, TPCN2 genes, TPCN1 genes, ULK1 genes, AKT2 genes, MTOR genes, CASP8 genes and/or LAMP2
Gene；
N2) using P1 as system used in the diagnosis of EBOV infection marks or auxiliary diagnosis EBOV infection method above-mentioned
A1 the application in) or a2)；
The P1 be TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3, MSR1,
MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2, MTOR, CASP8 and/or LAMP2 protein；
N3) using the G1 as EBOV infect mark in above-mentioned a1) or a2) in application；
N4) using the P1 as EBOV infect mark in above-mentioned a1) or a2) in application.
6. following W1) or product W2)：
W1 EBOV infection products) are treated and/or prevent, with G1 as target spot；
The G1 is TRAF2 genes, TP53 genes, CASP9 genes, RIPK3 genes, CASP7 genes, CIAPIN1 genes, TNF
Gene, BAX genes, CASP6 genes, TYRO3 genes, MSR1 genes, MARCO genes, HAVCR1 genes, IRS1 genes,
LRSAM1 genes, TPCN2 genes, TPCN1 genes, ULK1 genes, AKT2 genes, MTOR genes, CASP8 genes and/or LAMP2
Gene；
W2 EBOV infection products) are treated and/or prevent, with P1 as target spot；
The P1 be TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3, MSR1,
MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2, MTOR, CASP8 and/or LAMP2 protein.
7. following X1) or product X2)：
X1) diagnosis or auxiliary diagnosis EBOV infection product, are the system for detecting G1 expressions；
The G1 is TRAF2 genes, TP53 genes, CASP9 genes, RIPK3 genes, CASP7 genes, CIAPIN1 genes, TNF
Gene, BAX genes, CASP6 genes, TYRO3 genes, MSR1 genes, MARCO genes, HAVCR1 genes, IRS1 genes,
LRSAM1 genes, TPCN2 genes, TPCN1 genes, ULK1 genes, AKT2 genes, MTOR genes, CASP8 genes and/or LAMP2
Gene；
X2) diagnosis or auxiliary diagnosis EBOV infection product, are the system for detecting P1 contents；
The P1 be TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3, MSR1,
MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2, MTOR, CASP8 and/or LAMP2 protein.
8. according to arbitrary application or product described in claim 6 or 7 in claim 1-5, it is characterised in that：The G1 tables
Up to the expression that level is G1 described in blood；The P1 contents are the content of P1 described in blood.
The construction method of 9.EBOV infection relevant molecular characteristics spectrums, including：Respectively to EBOV infected groups and non-EBOV infected groups blood
RNA or protein in liquid carries out quantitative analyses, obtains EBOV virus loads relevant molecular characteristics spectrum according to analysis result；It is described
EBOV virus loads relevant molecular characteristics spectrum is G1 or P1；
The G1 is TRAF2 genes, TP53 genes, CASP9 genes, RIPK3 genes, CASP7 genes, CIAPIN1 genes, TNF
Gene, BAX genes, CASP6 genes, TYRO3 genes, MSR1 genes, MARCO genes, HAVCR1 genes, IRS1 genes,
LRSAM1 genes, TPCN2 genes, TPCN1 genes, ULK1 genes, AKT2 genes, MTOR genes, CASP8 genes and/or LAMP2
Gene；
The P1 be TRAF2, TP53, CASP9, RIPK3, CASP7, CIAPIN1, TNF, BAX, CASP6, TYRO3, MSR1,
MARCO, HAVCR1, IRS1, LRSAM1, TPCN2, TPCN1, ULK1, AKT2, MTOR, CASP8 and/or LAMP2 protein.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1323223A (en) * | 1998-10-09 | 2001-11-21 | 拜奥根有限公司 | Reversal of viral-induced systemic shock and respiratory distress by blockade of the lymphotoxin beta pathway |
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2016
- 2016-12-07 CN CN201611117710.8A patent/CN106591446A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1323223A (en) * | 1998-10-09 | 2001-11-21 | 拜奥根有限公司 | Reversal of viral-induced systemic shock and respiratory distress by blockade of the lymphotoxin beta pathway |
Non-Patent Citations (3)
| Title |
|---|
| ANITA K MCELROY等: "Biomarkers for understanding Ebola virus disease", 《BIOMARK MED》 * |
| JASON KINDRACHUK等: "Ebola Virus Modulates Transforming Growth Factor β Signaling and Cellular Markers of Mesenchyme-Like Transition in Hepatocytes", 《JOURNAL OF VIROLOGY》 * |
| S.BAIZE等: "Inflammatory responses in Ebola virus-infected patients", 《CLIN EXP IMMUNOL》 * |
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