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HAPLOIDENTICALSAMPLEANALYSIS VěraŠiffnerová
THEINSTITUTEOFHEMATOLOGYANDBLOODTRANSFUSION PRAGUE,CZECHREPUBLIC
• CuraDveapproachforpaDentswithhematologicalandimmunediseases • Increasingusage-improvementsinmedicaltreatment(cyclophosphamide) • Advantages
- Availabilityofdonor(parent,child,sibling)
- StrongGVLeffect
- Costsaving(comparedtoMUD) • Disadvantages
- GVHD(amajorcauseofmorbidityandmortality)
HAPLOIDENTICALTRANSPLANTATION
SolhMetal.Post-relapsesurvivalaWerhaploidenDcaltransplantaDonvsmatched-relatedormatched-unrelatedhematopoieDccelltransplantaDon.BoneMarrowTransplant.2016Jul;51(7):949-54. Dietrich,Setal.Post-transplantcyclophosphamide-basedhaplo-idenDcaltransplantaDonasalternaDvetomatchedsiblingorunrelateddonortransplantaDonfornon-Hodgkinlymphoma:aregistrystudybytheEuropeansocietyforbloodandmarrowtransplantaDon,Leukemia30,2086–2089(2016)
HAPLOIDENTICALTRANSPLANTATION
FrequentmechanismofanescapefromtheselecAvepressureofapaAent-specificGVLreacAon • Incidence-33%ofhaploidenDcalHSCT,AML• GeneDcrearrangement-absenceofthepaDent-specificHLAhaplotype+uniparentaldisomy• Occurslaterthanclassicalrelapse(12monthsx6months)• LowerageofpaDent(higherageprotecDve)• AcDvedisseaseattheDmeofHSCT,aGVHDandcGVHDbeforetherelapseonset
• Influenceonchoiceofsalvagetreatments• Donor lymphocyte infusion (DLI) - promoDon of rapid reconstrucDon of the immune system aWer
HSCTandresidualdiseasecontrolnoteffecDve,cansDmulatefurthergeneDcinstability
HLALOSSRELAPSE
• HLAHolotype • Twin 2.1.4 - analysis failed,
not possible to resolve 3-allelesample
• PaDent and donor HLAknown - possible to pre-selecttheexpectedresults?
HLALOSSRELAPSE-DETECTION
• Thelargesthematologyandhemato‑oncologycentreintheCzechRepublic • HaploidenDcalHSCTsince2014 • 67paDents(approx.¼ofatotalnumber)
• Donors-child(66%),sibling(20%),parent(14%)
INSTITUTEOFHEMATOLOGYANDBLOODTRANSFUSION
59% 12%
9%
7% 3% 4% 2% 2% 2%
acute myeloid leukemia
myelodysplastic syndrome
acute lymfoblastic leukemia and lymfoblastic lymphoma B-nonHodgkin lymphoma
myeloproliferative diseases
Overview of diagnoses
MATCH NUMBEROFPATIENTS 5/10 43 6/10 18 7/10 4 8/10 2
• HLAgroup • HLA typing of paDents and selected donors for transplant unit of adult (IHBT) and
paediatricpaDents(FacultyHospitalMotol,Prague) • Autoimmunedisorders&otherclinicalcondiDons • PharmacogeneDcsassociaDons
• Chimerismgroup
• Regularpost-transplantmonitoringsince1992 HLAmethods • NGS-firstchoicemethod,allpaDentsanddonors • SBT-confirmatorytyping,urgentpaDentanddonors • PCR-SSP-confirmatorytyping,urgentpaDentanddonors,diseaseassociaDons • qPCR-diseaseassociaDons,undergoingverificaDonprocessofkitsformedium-high
resoluDonHLAtyping
HLADEPARTMENT
• PaAent 1stsample-NGS 2ndsample-confirmatorytyping,SBT(aWerHSCTindicaDon)
• Relateddonors-siblings,parents,children,distantrelaAves
1stsample-NGS 10/10 HAPLOIDENTICAL+KIR
2ndsample(ifchosenlikeasuitabledonor)-confirmatorytyping,SBT(aWerHSCTindicaDon)
• Unrelateddonors
1sample-NGS+KIR KIR-relapseprotecDon,donorsdividedintocategoriesbasedonKIRB-content(neutral,beser,best) DPB1 - immunogenicity predicDon for mismatched donors (permissive, non-permissive HvG, non-permissiveGvH)
DONORSEARCH-STRATEGY
• Accreditedmethod -ISO15189-November2017 -EFI-undergoing,paperworksubmisedApril2018
• 1runperweek • 24samples/7loci(HLA-A,-B,-C,-DRB1,-DPB1,-DQA1and-DQB1) • HLAHolotypeV2 • HLATwin2.5.1
HLATYPING-NGS
• AdultpaAent+sibling-5/10matchedHSCT
• PredictedKIRB-contentgroupforprospecDvedonor1-Neutral • PredictedimmunogenicityoftheDPB1matchingforthispairis-Permissive • OtherrelaDvesnoavailable
CASEREPORT1-RAREALLELES
PATIENT DONOR-SIBLING
HLA-A *02:01 *02:01 *01:01 *02:01
HLA-B *15:01 *39:93 *08:01 *39:93
HLA-C *03:03 *12:03 *07:01 *12:03
HLA-DRB1 *11:03 *11:01 *08:04 *11:01
HLA-DQB1 *03:01 *03:01 *03:01 *03:01
HLA-DQA1 *05:05 *05:05 *05:05 *05:05
HLA-DPB1 *04:02 *01:01 *02:01 *01:01
• PaediatricpaAent+familystudy
Becameurgent PaDent-SBTinprocess,Sibling-PCR-SSP(ABDRcombiLR)
Nomatch+RarealleleB*15 ConfirmaDonneeded+NGSinprocess
HRfamilystudyresults
CASEREPORT2-RAREALLELES
PATIENT DONOR-SIBLING
HLA-A *01:01 *03:01 *02:01 *03:01
HLA-B *51:01 *27:05 *15:24 *07:02
HLA-C *14:02 *02:02 *03:03 *07:02
HLA-DRB1 *08:01 *04:01 *01:01 *01:01
HLA-DQB1 *04:02 *03:01 *05:01 *05:01
HLA-DQA1 *04:01 *03:03 *01:01 *01:01
HLA-DPB1 *04:02 *04:01 *14:01 *04:02
MOTHER FATHER
HLA-A *01:01 *03:01 *03:01 *02:01
HLA-B *51:01 *07:02 *27:05 *15:24
HLA-C *14:02 *07:02 *02:02 *03:03
HLA-DRB1 *08:01 *01:01 *04:01 *01:01
HLA-DQB1 *04:02 *05:01 *03:01 *05:01
HLA-DQA1 *04:01 *01:01 *03:03 *01:01
HLA-DPB1 *04:02 *04:02 *04:01 *14:01
• AdultpaAent+sibling-8/10matchedHSCT
• PredictedKIRB-contentgroupforprospecDvedonor1-Neutral • PredictedimmunogenicityoftheDPB1matchingforthispair-Permissive
CASEREPORT3-„ABNORMAL“MATCH
PATIENT DONOR-SIBLING
HLA-A *33:01 *68:01 *33:01 *68:01
HLA-B *14:02 *57:01 *14:02 *57:01
HLA-C *06:02 *08:02 *06:02 *08:02
HLA-DRB1 *15:01 *15:01 *15:01 *01:02
HLA-DQB1 *06:02 *06:02 *06:02 *05:01
HLA-DPB1 *04:01 *16:01 *04:01 *02:01
• AdultpaAent+3siblings-8/10matchedHSCT(10/10siblingnosuitable)
CASEREPORT4-„ABNORMAL“MATCH
PATIENT DONOR-SIBLING 10/10
DONOR-SIBLING 8/10
DONOR-SIBLING 5/10
HLA-A *02:01 *25:01 *02:01 *25:01 *32:01 *25:01 *02:01 *68:01
HLA-B *35:03 *18:01 *35:03 *18:01 *35:03 *18:01 *35:03 *08:01
HLA-C *04:01 *12:03 *04:01 *12:03 *04:01 *12:03 *04:01 *07:01
HLA-DRB1 *11:01 *12:01 *11:01 *12:01 *11:03 *12:01 *11:01 *03:01
HLA-DQB1 *03:01 *03:01 *03:01 *03:01 *03:01 *03:01 *03:01 *02:01
• AdultpaAent+familystudy-10/10(11/12)matchedHSCT
CASEREPORT5-RECOMBINATION
PATIENT DONOR-SIBLING 9/10
DONOR-SIBLING 1/10
HLA-A *24:02 *11:01 *24:02 *11:01 *03:01 *32:01
HLA-B *37:01 *39:06 *37:01 *39:06 *35:01 *40:02
HLA-C *06:02 *12:03 *06:02 *12:03 *04:01 *02:02
HLA-DRB1 *08:01 *16:01 *08:01 *16:01 *01:01 *11:01
HLA-DQB1 *04:02 *05:02 *04:02 *05:02 *05:01 *03:01
HLA-DQA1 *04:02 *01:02 *04:02 *01:02 *01:01 *05:05
HLA-DPB1 *01:01 *03:01 *01:01 *14:01 *03:01 *04:02
MOTHER FATHER
HLA-A *03:01 *11:01 *24:02 *32:01
HLA-B *35:01 *39:06 *37:01 *40:02
HLA-C *04:01 *12:03 *06:02 *02:02
HLA-DRB1 *01:01 *16:01 *08:01 *11:01
HLA-DQB1 *05:01 *05:02 *04:02 *03:01
HLA-DQA1 *01:01 *01:02 *04:02 *05:05
HLA-DPB1 *03:01 *14:01 *01:01 *04:02
SUMMARY • Haploidentical HSCT good variant of therapy
l no suitable fully matched donor is available l patient needs urgent HSCT
• Treatments now used for patients undergoing a haploidentical transplantation
(cyclophosphamide) have a positive impact on outcome measures (overall survival, disease-free progression, survival-free from acute or chronic graft-versus-host disease)
• Colaboration between laboratories • Searching for new markers and methods + Implementing in routine testing • Benefit for patients
ACKNOWLEDGEMENT
• HLAanalysisgroup MilenaVranáSaraNazarovaRenátaMacnerováŠárkaPůbalováHanaVondráčkováMilenaMelkováJanaNedvědováKateřinaŠimonová
• ChimerismgroupHanaČechová
• OmixonSupportTeam
P109 OVERVIEW OF POST-TRANSPLANT MONITORING IN PATIENTS AFTER HAPLOIDENTICAL ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION