Treatment of HIV and acute myeloid leukemia by allogeneic CCR5-d32 blood stem cell transplantation. Elena Knops
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1 Treatment of HIV and acute myeloid leukemia by allogeneic CCR5-d32 blood stem cell transplantation Elena Knops
2 Background - the Berlin patient so far the only person presumed to be cured from HIV by hematopoietic stem cell transplantation (HSCT) from a homozygous CCR5-d32 donor
3 Background - the Berlin patient so far the only person presumed to be cured from HIV by hematopoietic stem cell transplantation (HSCT) from a homozygous CCR5-d32 donor - failed attempts to reproduce cure by HSCT - viral rebound or death due to underlying malignancy
4 The whole story (drama in 5 acts) 48y old patient, alive, well and undetectable for HIV (RNA/DNA) over 4 years after allogeneic CCR5-d32 HSCT
5 The beginning (prologue) induction consolidation
6 The beginning (prologue) induction consolidation AML diagnosis in Jan 2011
7 The beginning (prologue) induction consolidation complete remission of AML
8 The beginning (prologue) induction consolidation to avoid interactions with chemotherapy TDF/FTC/RAL
9 Chapter I September 2012 female donor, 2 cycles high-dose cytarabine 10 out of 10 HLAmatched CCR5-d one week before SCT
10 Chapter I stem cells from female, 10/10 HLA-matched CCR5-d32 donor one week before SCT
11 Chapter I one week before SCT
12 Chapter I one week before SCT
13 Chapter II during transplant and until today on ART...
14 Chapter II June 2013 total of 8 courses of 5-azacytidine + 4 donor lymphocyte infusions...
15 Chapter III + CMV, HHV8, HSV <LOD (884250) in rectal and ileum biopsy
16 Chapter III + CMV, HHV8, HSV increased liver values <LOD (884250) in rectal and ileum biopsy
17 Chapter III + CMV, HHV8, HSV Tacrolimus + topical steroids <LOD (884250) in rectal and ileum biopsy
18 Chapter III + CMV, HHV8, HSV Tacrolimus + topical steroids <LOD (884250) in rectal and ileum biopsy
19 Chapter III + CMV, HHV8, HSV Tacrolimus + topical steroids... <LOD (884250) in rectal and ileum biopsy
20 Chapter III + CMV, HHV8, HSV Tacrolimus + topical steroids... - PBMCs - rectal + ileum - bone marrow - CSF
21 Chapter III Tacrolimus + topical steroids ABC/3TC/DTG <20 < <LOD (884250) in rectal and ileum biopsy <LOQ (15 Mio) 1/20 repl <LOQ (10 Mio) 1/10 repl <LOQ (10 Mio) <LOQ (3,8 Mio) 52 month after SCT
22 Chapter V month after SCT Mar Apr May Jun ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddpcr or qpcr, but positive signals in T-cell subsets: T CM : 0.2 Mio cells (ddpcr) 6.7cop/10 6 cells T EM : 0.36 Mio cells (qpcr) 5 cop/10 6 cells
23 Chapter V month after SCT Mar Apr May Jun ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddpcr or qpcr, but positive signals in T-cell subsets: T CM : 0.2 Mio cells (ddpcr) 6.7cop/10 6 cells T EM : 0.36 Mio cells (qpcr) 5 cop/10 6 cells G. Dunay, Hamburg: Ileum biopsy: 0.1 Mio cells measured: 1/4 replicates positive with LTR-, but negative with gag primers
24 Chapter V month after SCT Mar Apr May Jun ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddpcr or qpcr, but positive signals in T-cell subsets: T CM : 0.2 Mio cells (ddpcr) 6.7cop/10 6 cells T EM : 0.36 Mio cells (qpcr) 5 cop/10 6 cells G. Dunay, Hamburg: Ileum biopsy: 0.1 Mio cells measured: 1/4 replicates positive with LTR-, but negative with gag primers ICISTEM (J. Martinez-Picado & M. Salgado, Barcelona): qvoa: 23 Mio CD4 + T cells negative (IUPM < 0.031/10 6 CD4 T cells) Ileum biopsy: Mio CD45+ cells negative (<143/10 6 CD45+ cells) Rectum biopsy: Mio CD45+ cells negative (<34/10 6 CD45+ cells)
25 Chapter V month after SCT Mar Apr May Jun ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddpcr or qpcr, but positive signals in T-cell subsets: T CM : 0.2 Mio cells (ddpcr) 6.7cop/10 6 cells T EM : 0.36 Mio cells (qpcr) 5 cop/10 6 cells G. Dunay, Hamburg: Ileum biopsy: 0.1 Mio cells measured: 1/4 replicates positive with LTR-, but negative with gag primers ICISTEM (J. Martinez-Picado & M. Salgado, Barcelona): qvoa: 23 Mio CD4 + T cells negative (IUPM < 0.031/10 6 CD4 T cells) Ileum biopsy: Mio CD45+ cells negative (<143/10 6 CD45+ cells) Rectum I. biopsy: Hauber, Hamburg: Mio CD45+ cells negative (<34/10 6 CD45+ cells) mvoa: negative in plasma, negative in histology
26 Chapter V Rag#1DD hu-cd3 HIVp24Ag Spleen Liver LN I. Hauber, Hamburg: mvoa: negative in plasma, negative in histology
27 Chapter V PHENOTYPIC CO-RECEPTOR TROPISM ANALYSIS (sample before transplantation, PBMC) # sequencing genotypic prediction phenotypic analysis in Tcells Clone gp120-v3 amino acid sequence reads FPR (%) Magi (R5/X4) MT2 (X4) D1 CTRPNNNTREGIHIGPGRAFFTTGEIIGNIREASC 4 95,78 R5 R5 D2 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIKEAYC 2 95,64 R5 R5 D3 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIGEAYC 2 95,64 R5 R5 D4 CTRPNNNTRKGIHIGPGRAFFTTGEIIGNIREASC ,33 R5 R5 D5 CTRPNNNTRKGITIGPGRAFFTTGEIIGDIRQAHC ,67 R5 R5 D6 CTRPNNNTRKGIHIGSRKAFFTTGGIIGDIRQAYC 2 10,61 R5/X4 X4 D7 CTRPHTNTRKRIHIGPGRAFFTTGEIIGDIRQAYC 7 1,74 R5/X4 X4 D8 CTKPNNNTRKRIHIGPGRAFFTTGEIIGNIRQASC 2 1,74 R5 R5 D9 CTRPNNNIRKRIHIGPGRAFFTTGEIIGNIREAYC 3 1,16 R5/X4 X4 HxB2V3Bal control R5 51,8 R5 R5 HxB2 control X4 0 X4 X4 Monique Nijhuis, Utrecht, ICISTEM
28 Chapter V CO-RECEPTOR TROPISM (sample before transplantation, PBMC) # sequencing genotypic prediction phenotypic analysis in Tcells Clone gp120-v3 amino acid sequence reads FPR (%) Magi (R5/X4) MT2 (X4) D1 CTRPNNNTREGIHIGPGRAFFTTGEIIGNIREASC 4 95,78 R5 R5 D2 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIKEAYC 2 95,64 R5 R5 D3 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIGEAYC 2 95,64 R5 R5 D4 CTRPNNNTRKGIHIGPGRAFFTTGEIIGNIREASC ,33 R5 R5 D5 CTRPNNNTRKGITIGPGRAFFTTGEIIGDIRQAHC ,67 R5 R5 D6 CTRPNNNTRKGIHIGSRKAFFTTGGIIGDIRQAYC 2 10,61 R5/X4 X4 D7 CTRPHTNTRKRIHIGPGRAFFTTGEIIGDIRQAYC 7 1,74 R5/X4 X4 D8 CTKPNNNTRKRIHIGPGRAFFTTGEIIGNIRQASC 2 1,74 R5 R5 D9 CTRPNNNIRKRIHIGPGRAFFTTGEIIGNIREAYC 3 1,16 R5/X4 X4 HxB2V3Bal control R5 51,8 R5 R5 HxB2 control X4 0 X4 X4 Monique Nijhuis, Utrecht, ICISTEM
29 The end? (epilogue) More assays to be done: - qvoa + ultra microchimerism (ICISTEM: M. Salgado, Barcelona): lymph node (planned) - T-cell response assay (T. Harrer, Erlangen): PBMCs from donor (in progress) - proviral DNA load from further PBMC samples and lymph node (Institute of Virology, Cologne and others) - International collaboration to guide and investigate the potential for HIV cure by stem cell transplantation. The ICISTEM project is supported by amfar Research Consortium on HIV eradication (ARCHE) Research Grant # RGRL
30 Thanks to our group Institute of Virology, University of Cologne Rolf Kaiser Eva Heger Veronica Di Cristanziano Maria Neumann-Fraune Eugen Schülter Claudia Müller Saleta Sierra-Aragon Sascha Hasheminasab Institute for Immunogenetics, Kaiserslautern Martin Däumer, Alex Thielen University of Düsseldorf Hepatology & Infectiology Björn Jensen Falk Hüttig Dieter Häussinger Virology Nadine Lübke Haematology Guido Kobbe Moleculare Medizin Philipp Lang Junnat Hamdam Heinrich Pette Institut HH Ilona Hauber, Gabor Dunay, Johanna Eberhard, Silke Kummer, Julian Schulze zur Wiesch Med 3, University of Erlangen Thomas Harrer IciStem project Annemarie Wensing, Javier Martinez-Picado, Monique Nijhuis et al. MPI of Informatics, Saarbrücken Thomas Lengauer, Joachim Büch, Nico Pfeifer, Alejandro Pironti, Prabhav Kalaghatgi
31 Thanks to the Duesseldorf patient
32 Intermezzo gp160 gp120 p68 p55 gp41 p34 p25 p18 pos. control neg. control 02/2013 before SCT 06/ / / / /2016
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