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WO2014074501A1 - Marqueurs moléculaires permettant de diagnostiquer un rejet cellulaire aigu et une nécrose tubulaire aiguë - Google Patents

Marqueurs moléculaires permettant de diagnostiquer un rejet cellulaire aigu et une nécrose tubulaire aiguë Download PDF

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Publication number
WO2014074501A1
WO2014074501A1 PCT/US2013/068456 US2013068456W WO2014074501A1 WO 2014074501 A1 WO2014074501 A1 WO 2014074501A1 US 2013068456 W US2013068456 W US 2013068456W WO 2014074501 A1 WO2014074501 A1 WO 2014074501A1
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Prior art keywords
patient
sample
renal
rejection
suffering
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Stanley J. NAIDES
Marian MIKULA, Jr.
Feras M. ABU HANTASH
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Quest Diagnostics Investments LLC
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Quest Diagnostics Investments LLC
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/24—Immunology or allergic disorders
    • G01N2800/245—Transplantation related diseases, e.g. graft versus host disease
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/60—Complex ways of combining multiple protein biomarkers for diagnosis

Definitions

  • the present disclosure generally relates to methods for diagnosis of renal graft rejections including, in particular, acute cellular rejection (ACR), acute tubular necrosis (ATN), in a renal transplant recipient. Once the rejection is detected, methods for treating the rejection are provided.
  • ACR acute cellular rejection
  • ATN acute tubular necrosis
  • Renal transplant recipients may suffer graft rejections, such as acute cellular rejection (ACR), acute tubular necrosis (ATN), antibody mediated rejection (ABMR), and other types of rejection reactions.
  • ACR acute cellular rejection
  • ATN acute tubular necrosis
  • ABMR antibody mediated rejection
  • T-cells part of the adaptive immune system, play a role in ACR.
  • Natural killer cells (NKC) of the innate immune system are known to play a role in ATN and ABMR.
  • Renal graft rejections including ACR and ATN, are usually diagnosed on the basis of histologic evaluation performed in response to biochemical evidence of graft impairment (e.g., elevated creatinine levels). There is a need to develop more accurate and noninvasive tests for renal graft rejections that may allow earlier detection and intervention.
  • the present disclosure provides a method for identifying a renal transplant patient as likely suffering renal graft rejection, comprising: determining the expression level of one or more genes selected from the group consisting of c-abl oncogene 1 non-receptor tyrosine kinase (ABL1), CD 160 molecule (NK1 or CD 160), natural cytotoxicity triggering receptor 3 (NK3 or NCR3), granzyme B (GZMB), perforin 1 (PRF1), forkhead box P3 (FOXP3) and chemokine (C-X-C motif) ligand 10 (CXCL10) in a sample isolated from a patient; and identifying the patient as likely suffering renal graft rejection if at least one of the one or more genes is overexpressed in the sample.
  • ABL1 c-abl oncogene 1 non-receptor tyrosine kinase
  • NK1 or CD 160 CD 160 molecule
  • NK3 or NCR3 natural cytotoxicity triggering receptor 3
  • the renal graft rejection comprises one or more conditions selected from the group consisting of acute cellular rejection (ACR), acute tubular necrosis (ATN), antibody mediated rejection (ABMR), chronic cellular rejection, borderline rejection, diabetic nephropathy, transplant nephropathy, and minimal interstitial fibrosis or tubular atrophy.
  • ACR acute cellular rejection
  • ATN acute tubular necrosis
  • ABMR antibody mediated rejection
  • chronic cellular rejection borderline rejection
  • diabetic nephropathy transplant nephropathy
  • minimal interstitial fibrosis or tubular atrophy the patient is identified as likely suffering ATN or ACR if at least one of CXCLIO, FOXP3, GZMB or PRFl is overexpressed in the sample, and in particular, is identified as likely suffering ATN or ACR if at least two of CXCLIO, FOXP3, GZMB or PRFl are overexpressed in the sample.
  • the patient is identified as likely suffering ATN if (a) ABL1
  • Overexpression is determined as an increase of expression level above a reference range of expression level of the corresponding gene in a normal population, and the reference ranges are provided in Table 1.
  • FIG. 1A-B show transcription levels of examined genes (GZMB, PRFl , FOXP3 and CXCLIO) relative to ABL1 (FIG. 1A) and CD3e (FIG. IB) in renal transplant recipients.
  • Patient classification is based on renal histology. "Other” includes chronic cellular rejection, borderline rejection, diabetic nephropathy, transplant nephropathy, and minimal interstitial fibrosis or tubular atrophy.
  • X-axis shows patient numbers. Error bars are 95% CI based on 4 replicates. Bars without errors bars exceed the Y-axis range. Reference ranges are in Table 1.
  • FIG. 2A-B show transcription levels of natural killer genes (NK1 and NK2) relative to ABL1 (FIG. 2A) and CD3e (FIG. 2B) in renal transplant recipients.
  • Classification is based on renal histology. "Other” includes chronic cellular rejection, borderline rejection, diabetic nephropathy, transplant nephropathy, and minimal interstitial fibrosis or tubular atrophy.
  • X-axis shows patient numbers. Error bars are 95% CI based on 4 replicates. Bars without errors bars exceed the Y-axis range. Reference ranges are in Table 1.
  • FIG. 3A-M present the relative gene expression quantities of the marker genes (as indicated) for a patient, S01, that did not experience renal graft rejections.
  • the x-axis indicates days following the renal transplant.
  • the dotted lines show the calculated elevation thresholds for each gene.
  • FIG. 3A-F use Abll for normalization (as indicated) and FIG. 3G- M use CD3d for normalization (as indicated).
  • FIG. 4A-W present relative gene expression quantities of the marker genes (as indicated) for a patient, R01, that experienced acute cellular rejection (ACR).
  • the x-axis indicates days following the renal transplant.
  • the dotted lines show the calculated elevation thresholds for each gene.
  • FIG. 4A-J use Abll for normalization (as indicated) and FIG. 4K- W use CD3d for normalization (as indicated).
  • the gene expression levels for some of the genes are twice, at different scales for clarity (see, e.g., FIG. 4A-B).
  • the present disclosures are based on the identification of gene expression markers useful for detection of renal graft rejections in a renal transplant patient.
  • nucleic acid includes a combination of two or more nucleic acids, and the like.
  • One embodiment of the present disclosure provides a method for identifying a renal transplant patient as likely suffering renal graft rejection.
  • the method entails determining the expression level of one or more genes selected from the group consisting of c-abl oncogene 1 non-receptor tyrosine kinase (ABL1), CD 160 molecule (NK1 or CD 160), natural cytotoxicity triggering receptor 3 (NK3), granzyme B (GZMB), perforin 1 (PRF1), forkhead box P3 (FOXP3) and chemokine (C-X-C motif) ligand 10 (CXCL10 or IP 10) in a sample isolated from a patient.
  • ABL1 c-abl oncogene 1 non-receptor tyrosine kinase
  • NK1 or CD 160 CD 160 molecule
  • NK3 natural cytotoxicity triggering receptor 3
  • GZMB granzyme B
  • PRF1 perforin 1
  • FOXP3 forkhead box P
  • the expression levels of all of the genes are generally within a "reference range” in samples isolated from patients not suffering renal graft rejections ("normal" patients). The majority of patients that suffer from one or more of such rejections, by contrast, exhibited "overexpression" of one or more of these genes.
  • the expression level of a gene in a sample in one aspect, is expressed as a "relative quantity" or "RQ" number, which is the ratio between the measured expression of the gene in a test sample and the measured expression of the gene in a control sample.
  • the control sample is a sample isolated from a patient that does not suffer renal graft rejections.
  • the control sample is a sample prepared by pooling two or more such control samples.
  • a virtual control sample can be prepared by mathematically summarizing the expression of the gene in more than one control samples.
  • a gene is "overexpressed" in a test sample when the relative quantity of the gene in the test sample is above a "reference range.”
  • a “reference range” reflects a normal range of relative quantities that a gene exhibits in control samples.
  • a gene's reference range can be determined by measuring the expression levels of the gene in two, or three or more control samples, following statistical summarization of the expression levels. For instance, Gaussian distribution can be used to calculate the reference ranges.
  • the raw gene expression value can be normalized against a control gene.
  • the control gene is a housekeeping gene.
  • the control gene is ABL1
  • the control gene is CD3e molecule epsilon (CD3e). Exemplary reference ranges for genes in the present disclosure are provided in Table 1.
  • Renal graft rejections refer to rejections of a transplanted renal tissue by the recipient's immune system, which cause injury in or destroy the transplanted renal tissue.
  • Various types of renal graft rejections have been observed clinically, including without limitation acute cellular rejection (ACR), acute tubular necrosis (ATN), antibody mediated rejection (ABMR), chronic cellular rejection, borderline rejection, diabetic nephropathy, transplant nephropathy, and minimal interstitial fibrosis or tubular atrophy.
  • ACR acute cellular rejection
  • ATN acute tubular necrosis
  • ABMR antibody mediated rejection
  • Chronic cellular rejection borderline rejection
  • diabetic nephropathy transplant nephropathy
  • transplant nephropathy transplant nephropathy
  • minimal interstitial fibrosis or tubular atrophy minimal interstitial fibrosis or tubular atrophy.
  • the patient from which the sample is isolated can be identified as likely suffering from one or more of renal graft rejections. This is based on the fact that none of the normal patients exhibited overexpression of any of these genes, and only those that suffer from the rejection did.
  • the experimental example demonstrates, when at least one of GZMB, PRFl, FOXP3 or CXCLIO is overexpressed, the patient likely suffers ATN or ACR.
  • the patient when at least two, or alternatively at least three, or all four of GZMB, PRFl, FOXP3 or CXCLIO are overexpressed, the patient likely suffers ATN or ACR.
  • the present disclosure further provides markers for distinguishing ATN from ACR. It has been discovered, unexpectedly, that ABLl, a putative housekeeping gene, was overexpressed in all ATN patients. Furthermore, many ATN patients exhibited
  • the patient is identified as likely suffering ATN if (a) ABLl and (b) NKl or NK3 are overexpressed in the sample.
  • the patient is identified as likely suffering ACR if none of ABLl, NKl and NK3 is overexpressed in the sample.
  • the method of the present disclosure entails the detection of the gene expression levels of the genes in sample that is isolated before the rise of the creatinine level in a patient, or at a time that is prior to the anticipated rise of the creatinine level.
  • a method for identifying a renal transplant patient as likely suffering renal graft rejection comprising determining the expression level of one or more genes selected from the group consisting of c-abl oncogene 1 non-receptor tyrosine kinase (ABLl), CD 160 molecule (NKl), natural cytotoxicity triggering receptor 3 (NK3), granzyme B (GZMB), perforin 1 (PRFl), forkhead box P3 (FOXP3) and chemokine (C-X-C motif) ligand 10 (CXCLIO) in a sample isolated from a patient that has undergone renal transplant and has a level of creatinine within a normal physiological range; and identifying the patient as likely suffering renal graft rejection if at least one, or two, or three, or four of the one or more genes are overexpressed in the sample.
  • ABLl c-abl oncogene 1 non-receptor tyrosine kinase
  • NKl CD 160 molecule
  • Normal physiological ranges are known the art. For instance, normal levels of creatinine in the blood are approximately 0.6 to 1.2 milligrams (mg) per deciliter (dL) in adult males and 0.5 to 1.1 milligrams per deciliter in adult females. Therefore, in some embodiments, the patient undergoing the detection has a blood creatinine level that is equal to or lower than about 1.0 milligrams per deciliter, 1.1 milligrams per deciliter, or 1.2 milligrams per deciliter, or 1.3 milligrams per deciliter, or 1.4 milligrams per deciliter, or 1.5 milligrams per deciliter, or 2 milligrams per deciliter.
  • the expression levels of one or more genes are determined in a sample isolated from a renal transplant patient.
  • sample refers to any liquid or solid material containing nucleic acids.
  • a sample is a blood or tissue sample from a human patient.
  • a sample comprises peripheral blood or plasma.
  • the expression level of a gene refers to the mRNA expression level of the gene.
  • Various methods of nucleic acid extraction are suitable for isolating total RNA or mRNA. See Maniatis et al., Molecular Cloning, A Laboratory Manual, 2d, Cold Spring Harbor Laboratory Press, page 16.54 (1989). Numerous commercial kits also yield suitable RNA including, but not limited to, QIAampTM mini blood kit, Agencourt GenfindTM, Roche Cobas® Roche MagNA Pure® or phenolxhloroform extraction using Eppendorf Phase Lock Gels®, and the NucliSens extraction kit (Biomerieux, Marcy l'Etoile, France). In other methods, mRNA may be extracted from patient blood/bone marrow samples using MagNA Pure LC mRNA HS kit and Mag NA Pure LC Instrument (Roche Diagnostics Corporation, Roche Applied Science, Indianapolis, IN).
  • Nucleic acid extracted from tissues, cells, plasma or serum can be amplified using nucleic acid amplification techniques well known in the art. Many of these amplification methods can also be used to detect the presence of mutations simply by designing
  • oligonucleotide primers or probes to interact with or hybridize to a particular target sequence in a specific manner.
  • these techniques can include the polymerase chain reaction (PCR) reverse transcriptase polymerase chain reaction (RT-PCR), nested PCR, ligase chain reaction.
  • PCR polymerase chain reaction
  • RT-PCR reverse transcriptase polymerase chain reaction
  • nested PCR ligase chain reaction
  • RNA reporters S., et al, AIDS 7 (suppl 2):S11-S 14, (1993), amplifiable RNA reporters, Q-beta replication, transcription- based amplification, boomerang DNA amplification, strand displacement activation, cycling probe technology, isothermal nucleic acid sequence based amplification (NASBA). See Kievits, T. et al, J Virological Methods 35:273-286, (1991), Invader Technology, or other sequence replication assays or signal amplification assays.
  • NASBA isothermal nucleic acid sequence based amplification
  • RNA to cDNA Some methods employ reverse transcription of RNA to cDNA. As noted, the method of reverse transcription and amplification may be performed by previously published or recommended procedures, which referenced publications are incorporated herein by reference in their entirety.
  • Various reverse transcriptases may be used, including, but not limited to, MMLV RT, RNase H mutants of MMLV RT such as Superscript and Superscript II (Life Technologies, GIBCO BRL, Gaithersburg, Md.), AMV RT, and thermostable reverse transcriptase from Thermus thermophilus .
  • RNA extracted from plasma or serum is the protocol adapted from the Superscript II Preamplification system (Life Technologies, GIBCO BRL, Gaithersburg, Md.; catalog no. 18089-011), as described by Rashtchian, A., PCR Methods Applic. 4:S83-S91, (1994), adapted as follows.
  • RNA extracted from plasma or serum in 13 ⁇ of DEPC-treated water is added to a clean microcentrifuge tube. Then one microliter of either oligo (dT) (0.5 mg/ml) or random hexamer solution (50 ng/ ⁇ ) is added and mixed gently. The mixture is then heated to 70 degrees centigrade for 10 minutes and then incubated on ice for one minute.
  • oligo 0.5 mg/ml
  • random hexamer solution 50 ng/ ⁇
  • the reaction is then terminated by incubating the tube at 70°C for 15 minutes, and then placing it on ice.
  • the reaction is collected by brief centrifugation, and 1 ⁇ of RNase H (2 units) is added followed by incubation at 37°C for 20 minutes before proceeding to nucleic acid amplification.
  • To the cDNA mixture add the following: 8 ⁇ of 10* synthesis buffer (200 mM Tris- HC1, pH 8.4, 500 mM KC1, 25 mM magnesium chloride, 1 mg/ml of BSA), 68 ⁇ sterile double-distilled water, 1 ⁇ amplification primer 1 (10 ⁇ ), 1 ⁇ amplification primer 2 (10 ⁇ ), 1 ⁇ Taq DNA polymerase (2-5 U/ ⁇ ). Mix gently and overlay the reaction mixture with mineral oil. The mixture is heated to 94°C for 5 minutes to denature remaining
  • PCR amplification is then performed in an automated thermal-cycler for 15-50 cycles, at 94°C for 1 minute, 55° for 30 to 90 seconds, and 72°C for 2 minutes.
  • the present disclosure further provides methods for treating the rejection.
  • the terms “treating” or “treatment” or “alleviation” refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) the targeted pathologic condition or disorder.
  • a subject is successfully “treated” for a disorder if, after receiving a therapeutic agent according to the methods of the present disclosure, the subject shows observable and/or measurable reduction in or absence of one or more signs and symptoms of a particular disease or condition.
  • Methods for treating renal graft rejections are known in the art.
  • Acute tubular necrosis (ATN) can be treated with dialysis or plasma exchange. See, e.g., Esson and Schrler, Ann. Intern. Med. 137:744-52 (2002).
  • ACR acute cellular rejection
  • lymphocyte-depleting antibodies or OKT3 can be administered.
  • Plasma exchange can also be useful for treating or alleviating symptoms of ACR.
  • This example investigated differential expression patterns of several genes in patients with various forms of renal graft failure. These genes included PRF1, GZMB, FOXP3, and CXCL10 (IP 10). Initial experiments used CD3e and ABLl (a housekeeping gene) as internal controls for relative quantitation.
  • this example observed aberrant levels of ABLl expression (relative to CD3e) in samples from ATN patients. Since natural killer (NK) cells produce ABLl but not CD3e, this example also investigated whether two known markers of NK cells, NK1 and NK3, might help further differentiate ATN from ACR.
  • NK natural killer
  • FIG. 1-2 show the distribution of histology. One hundred twenty apparently healthy, ambulatory, community-dwelling adults were recruited to establish reference ranges. Individuals with a known history of renal, autoimmune, or inflammatory disease were excluded.
  • PAXgene® tubes QIAGEN Inc, Valencia, CA
  • Roche MagNA Pure method Roche Applied Science, Indianapolis, IN
  • Extracted RNA was then reverse-transcribed to generate cDNA for real- time PCR on the ABI Viia® 7 thermal cycler (Applied Biosystems, Foster City, CA).
  • Targeted gene expression markers included two cytotoxic T lymphocyte markers (GZMB; PRF1), the regulatory T lymphocyte transcription factor FOXP3 (FOXP3), a chemokine expressed in T lymphocytes and monocytes (IP 10, CXCLIO); and the NK cell markers NKl and NK3.
  • GZMB cytotoxic T lymphocyte markers
  • FOXP3 regulatory T lymphocyte transcription factor 3
  • IP 10, CXCLIO a chemokine expressed in T lymphocytes and monocytes
  • NKl and NK3 included two cytotoxic T lymphocyte markers (GZMB; PRF1), the regulatory T lymphocyte transcription factor FOXP3 (FOXP3), a chemokine expressed in T lymphocytes and monocytes (IP 10, CXCLIO); and the NK cell markers NKl and NK3.
  • the T lymphocyte marker CD3e and the pan-cell marker ABLl were used as constitutive ly expressed endogenous normalizer genes.
  • Cycle threshold (Ct) values were compared to a normal pooled control sample via the delta-delta-Ct method to give a relative quantity (RQ) for each transcript (normalized to one of the marker genes). Markers with RQ values above the reference range values were considered elevated.
  • ACR For ACR, when the expression levels of the genes were evaluated using ABLl as control: 4 of 5 ACR patients showed markedly elevated CXCLIO; 1 showed elevated GZMB; and 1 had slightly elevated PRF1. One ACR patient had elevated NK3, but all other ACR patients had normal levels of both NK markers.
  • CD3e 3 of 5 ACR patients showed markedly elevated CXCL10; 1 had elevated GZMB; 1 had slightly elevated PRF1; 1 had elevated ABL1; and none had elevated FOXP3.
  • NK cell mRNAs for NK1 and NK3 have potential value as markers of ATN in renal transplant recipients.
  • information from markers of ACR CXCL10, FOXP3, GZMB, PRF1 and NK cells (NK1, NK3) may help distinguish between ACR and ATN.
  • Both ATN and ACR may be associated with elevated levels of more than one ACR markers.
  • ATN is characterized by elevated ABL1 plus NK1 or NK3, along with more than one ACR marker, and ACR is generally characterized by normal levels of ABL1, NK1, and NK3.
  • FIG. 3A-M present the relative quantity of the genes measured from one of the stable patients, R01.
  • the marker genes CXCL10 (FIG. 3 A), FOXP3 (FIG. 3B), GZMB (FIG. 3C), NK1 (FIG. 3D), NK3 (FIG. 3E) and PRF1 (FIG. 3F)
  • FIG. 3G-M present the relative quantity of the genes measured from one of the stable patients, R01.
  • the marker genes CXCL10 (FIG. 3 A), FOXP3 (FIG. 3B), GZMB (FIG. 3C), NK1 (FIG. 3D), NK3 (FIG. 3E) and PRF1 (FIG. 3F)
  • FIG. 3G-M present the relative quantity of the genes measured from one of the stable patients, R01.
  • the marker genes CXCL10 (FIG. 3 A), FOXP3 (FIG. 3B), GZMB (FIG. 3C), NK1 (FIG. 3
  • FIG. 4A-W The gene expression data from one of the two ACR patients, R01, are presented in FIG. 4A-W.
  • CXCL10 FIG. 4A-B
  • GZMB FIG. 4D-E
  • NK1 FIG. 4F-G
  • PRF1 FIG. 41- J
  • NK3 FIG. 4H
  • FIG. 4H Only FOXP3 (FIG. 4C) did not show elevation. Similar elevation of gene expression levels is shown in FIG. 4K-V, when normalized with CD3e. Again, only FOXP3 (FIG. 4M) did not show elevation and the elevation for other genes were seen at as early as 7 weeks post-transplantation (FIG. 4K-L). The elevation of these gene markers, therefore, are valid markers predicting the renal rejections. It was also observed that the elevation started even before the rise of creatinine, a conventional clinical marker for renal rejections.

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Abstract

La présente invention concerne des méthodes permettant d'identifier un patient souffrant probablement d'un rejet de greffe de rein, lesdites méthodes comprenant de déterminer le niveau d'expression d'un ou plusieurs gènes sélectionnés dans le groupe comprenant ABL1, NK1, NK3, GZMB, PRF1, FOXP3 et CXCL10 dans un échantillon isolé d'un patient et d'identifier le patient comme souffrant probablement d'un rejet de greffe de rein si au moins un du ou des gènes est surexprimé dans l'échantillon. La présente invention concerne également des méthodes permettant de traiter le rejet lorsqu'un patient souffrant d'un rejet de greffe de rein est identifié.
PCT/US2013/068456 2012-11-06 2013-11-05 Marqueurs moléculaires permettant de diagnostiquer un rejet cellulaire aigu et une nécrose tubulaire aiguë Ceased WO2014074501A1 (fr)

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Cited By (9)

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WO2016097259A1 (fr) * 2014-12-19 2016-06-23 INSERM (Institut National de la Santé et de la Recherche Médicale) Procédés de prédiction d'altérations de greffes
US9752191B2 (en) 2009-07-09 2017-09-05 The Scripps Research Institute Gene expression profiles associated with chronic allograft nephropathy
US10443100B2 (en) 2014-05-22 2019-10-15 The Scripps Research Institute Gene expression profiles associated with sub-clinical kidney transplant rejection
WO2020148358A1 (fr) * 2019-01-17 2020-07-23 Katholieke Universiteit Leuven Biomarqueurs à base d'arnm pour rejet de greffe à médiation par anticorps
EP3845665A1 (fr) * 2016-08-17 2021-07-07 The Regents Of The University Of California Nouvelle méthode basée sur une immunosonde, permettant d'évaluer un état de lésion organique par un dosage d'adn acellulaire (cfdna) à base de biofluide
US11104951B2 (en) 2014-05-22 2021-08-31 The Scripps Research Institute Molecular signatures for distinguishing liver transplant rejections or injuries
EP3764877A4 (fr) * 2018-03-12 2021-12-15 The Regents of The University of California Évaluation du statut de rejet de greffe par analyse de la diversité de répertoire de sous-unités de récepteur de lymphocytes t
US12329365B2 (en) 2020-12-17 2025-06-17 Kidneymetrix Inc. Kits for stabilization of urine samples at room temperature
US12404547B1 (en) 2014-03-14 2025-09-02 Caredx, Inc. Methods and systems for monitoring a recipient of an allograft

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US12404547B1 (en) 2014-03-14 2025-09-02 Caredx, Inc. Methods and systems for monitoring a recipient of an allograft
US10443100B2 (en) 2014-05-22 2019-10-15 The Scripps Research Institute Gene expression profiles associated with sub-clinical kidney transplant rejection
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US11926868B2 (en) 2016-08-17 2024-03-12 The Regents Of The University Of California Immunoprobe-based method to assess organ injury status through a biofluid-based cell-free DNA (cfDNA) assay
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EP3764877A4 (fr) * 2018-03-12 2021-12-15 The Regents of The University of California Évaluation du statut de rejet de greffe par analyse de la diversité de répertoire de sous-unités de récepteur de lymphocytes t
US12351874B2 (en) 2018-03-12 2025-07-08 The Regents Of The University Of California Assessing transplant rejection status by analysis of t-cell receptor subunit repertoire diversity
US20220081715A1 (en) * 2019-01-17 2022-03-17 Katholieke Universiteit Leuven mRNA-BASED BIOMARKERS FOR ANTIBODY-MEDIATED TRANSPLANT REJECTION
WO2020148358A1 (fr) * 2019-01-17 2020-07-23 Katholieke Universiteit Leuven Biomarqueurs à base d'arnm pour rejet de greffe à médiation par anticorps
US12329365B2 (en) 2020-12-17 2025-06-17 Kidneymetrix Inc. Kits for stabilization of urine samples at room temperature

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