WWW.UHKT.CZ

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