Metabolomics Profiling after Allogeneic Hematopoietic Stem Cell Transplantation Unravels a Specific Signature in Human Acute GVHD

Abstract Introduction Many efforts have been attempted to improve our understanding of GVHD pathophysiology but few focused on human GVHD. Most studies have focused on the central role played by immune cells. However, recent researches have highlighted the influence of tissue microenvironment or of host microbiota in regulating allo-immune response. In human, circulating metabolites are produced by both tissues and gut microbiota. Among them, many biochemical compounds regulate immune cells function and could be the bridge between microbiota, tissues and the immune system. The aim of this study was to characterize metabolomics alterations associated with allo-HSCT and to determine biochemical pathway specifically involved during acute GVHD. Methods We collected donors and recipients' frozen plasma from a monocentric (n=43) and a multicentric cohort (n=56) of patients who underwent transplantation from an HLA-identical sibling in France. Donors' samples were collected before stem cell collection. Recipient's samples were obtained at acute GVHD onset or at day + 90 in patients without GVHD. After protein removal, plasma extracts were analyzed by two separate reverse phase (RP)/UPLC-MS/MS methods with positive ion mode electrospray ionization (ESI), one for analysis by RP/UPLC-MS/MS with negative ion mode ESI and one for analysis by HILIC/UPLC-MS/MS with negative ion mode ESI. Compounds were identified by comparison to library entries of purified standards or recurrent unknown entities. Peaks were quantified using area-under-the-curve. All statistical analyses were performed with R 3.5.0 and the MetaboAnalystR package. Results More than 800 circulating metabolites were identified in the 2 cohorts, belonging to lipid (43.6%), amino acid (22.2%), xenobiotics (19.1%), nucleotide (4.2%), carbohydrate (3.6%), cofactors and vitamins (3.4%), peptide (2.8%) and energy (1.1%) pathways. After allo-HSCT, the comparison of metabolomics profiles of recipients without GVHD with those of healthy related donors revealed major changes in 222 metabolites (with a significance threshold at p<0.05). SAM and Volcano Plot analysis (with a fold change (FC) > 2 and p<0.05) highlighted significant modification in 43 metabolites. In the absence of acute GvHD, allo-HSCT was mainly characterized by a higher level of primary bile acids, mono and diacylglycerol, but a decrease of phospholipid, sphingolipid and secondary bile acid metabolites. Interestingly, polyamine, including N-acetyl putrescine (FC=5.5, p<0.001) and N-acetyl spermidine (FC=3.7, p<0.0001), were increased, suggesting that these microbiota-derived metabolites might play a protective role on gut integrity in patients without GVHD. After comparison of recipients with or without GvHD, we were able to identify specific metabolomics changes associated with the onset of GvHD, irrespective of age, sex and feeding of patients. Acute GvHD was associated with a major decrease of plasmalogen and lysoplasmalogen (FC<0.5, p<0.0001), that may play a role in protection against oxidative stress induced by ROS. By contrast, medium and long chain fatty and polyunsaturated acid were strongly increased (FC>2, p<0.01). Most metabolites of amino acid pathways were decreased at onset of acute GvHD. Among them, Aryl Hydrocarbon receptor ligands were diminished in patients with GvHD, especially host- and microbiota-derived tryptophan metabolites such as 3-indoxyl sulfate (p=0.0001), indole acetate (p=0.03), indole propionate (p=0.03) and N-acetyl kynurenine (p=0.04). Many other gut microbiota-derived metabolites were significantly decreased in patients with GvHD, suggesting that major microbiota injury after allo-HSCT may regulate allo-immune responses through the production of metabolites with immunomodulatory properties. Discussion Our results demonstrate that allo-HSCT is associated with major metabolomics changes in recipients, which might me due to drug intakes, metabolic stress or microbiota alteration. GvHD was characterized by specific changes in complex lipid metabolism and amino acid metabolites that are involved in immune cells regulation and inflammation. This study highlights the potential role of circulating metabolites in GvHD pathophysiology that could be targeted for prophylaxis or treatment. Figure Figure. Disclosures Peffault De Latour: Pfizer Inc.: Consultancy, Honoraria, Research Funding; Alexion Pharmaceuticals, Inc.: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Research Funding; Amgen Inc.: Research Funding.

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Impact of Antithymocyte Globulin-Containing Conditioning Regimens on Patients Undergoing Allogeneic Stem Cell Transplantation for Progressive Myelodysplastic Syndrome: A Report From the SFGM-TC Group

Blood ◽

10.1182/blood.v118.21.3035.3035 ◽

2011 ◽

Vol 118 (21) ◽

pp. 3035-3035

Author(s):

Ibrahim Yakoub-Agha ◽

Gandhi Damaj ◽

Marie Robin ◽

Stephane Vigouroux ◽

Alice Garnier ◽

...

Keyword(s):

Stem Cell ◽

Stem Cell Transplantation ◽

Board Of Directors ◽

Cumulative Incidence ◽

Research Funding ◽

Acute Gvhd ◽

Conditioning Regimen ◽

Refractory Disease ◽

Advisory Committees ◽

Grade 3

Abstract Abstract 3035 Background: due to a risk of relapse of underlying disease in patients transplanted with progressive malignancy, the use of antithymocyte globulins (ATG), incorporated within the conditioning regimen prior to allogeneic stem cell transplantation (allo-SCT), is still controversial. We report here on a study of 245 consecutive patients transplanted between January 1999 and December 2009 in 26 French and Belgian centers for progressive MDS, defined as stable, untreated, relapsed or refractory disease. Patients and Methods: Inclusion criteria included patients aged over 18 who received allo-SCT from either a sibling (n=153) or HLA-A, -B, -C, -DRB1 and -DQB1 allele matched unrelated donor (10/10) (n=86) for MDS or AML/RAEB-t (with 20–30% BM blasts). Data quality was ensured using computerized discrepancy errors and vigorous on-site data verification of every single file. A qualified research technicien has been appointed by the University-Hospital of Lille to assist on-site centers that couldn't meet data quality requirements. HLA matching was double-checked by the French Bone Marrow Donor Registry. Results: The first 239 files analyzed until now are presented, including 154 males and 85 females. According to the WHO classification at diagnosis, 85 patients had RA/RARS/RCMD, 86 RAEB1, 62 REAB2 and 6 RAEB-t/AML. Sixty-six patients had progressed to a more advanced disease before allo-SCT. At diagnosis, 102 patients had an IPSS int-2 or higher. Cytogenetic IPSS was recorded as favorable (n=109), intermediate (n=61), unfavorable (n=63) and missing (n=6). Disease status at transplant was established as follows: relapsed or refractory disease (n=106) and untreated or stable disease without hematological improvement (n=133). Median age at transplantation was 53 years (range, 20–70). Patients received myeloablative conditioning (n=105) and nonmyeloablative (n=134) including busulfan-based regimens (n=127), TBI-based regimens (n=92) or other alkylating-agent-based regimens (n=20). In this series, 95 patients (40%) received ATG as part of conditioning ('ATG' group), whereas 144 did not ('no-ATG' group). The analysis reference date of April 1st 2011, median follow-up in survivors was 50 months (IQR, 33–92) with 59 patients having died of relapse and 77 of TRM. The estimated 3-year OS and EFS was respectively 42.3%, and 32.4%. The probability of relapse, overall and event-free survival at 3 years was not significantly different between the two groups. In contrast, the cumulative incidence of grade 2–4 acute GVHD was 48% in the no-ATG group and 30% ATG group (P <.001) and the cumulative incidence of grade 3–4 acute GVHD was 24% and 11% respectively (P <.001). Although the cumulative incidence of chronic GVHD was similar in the no-ATG and ATG groups (64% vs 46%, p=.15), a trend for a lower TRM was observed in the ATG group (22% vs 31%, p=.06). In multivariate analysis, the absence of use of ATG was the strongest parameter associated with an increased risk of acute grade 2–4 [HR = 2.28, 95% CI: 1.39–3.74, p=.001] and grade 3–4 GVHD [HR = 2.19, 95% CI: 1.04–4.61, p=.035]. In conclusion, the addition of ATG to the conditioning regimen resulted in a decreased incidence of acute GVHD without increasing relapse rates and compromising patient survival undergoing allo-SCT for progressive MDS. Disclosures: Yakoub-Agha: Genzyme: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Honoraria, Research Funding; Fresinus: Honoraria, Membership on an entity's Board of Directors or advisory committees; Roche: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria. Michallet:Genzyme: Honoraria, Membership on an entity's Board of Directors or advisory committees; Fresinus: Honoraria, Membership on an entity's Board of Directors or advisory committees. Deconinck:Celgene: Honoraria. Mohty:Genzyme: Honoraria, Membership on an entity's Board of Directors or advisory committees.

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Graft-Versus-Host Disease Following Myeloablative Busulfan Plus Fludarabine and Busulfan Plus Cyclophosphamide For Allogeneic Hematopoietic Stem Cell Transplantation In Acute Myeloid Leukemia In First Complete Remission

Blood ◽

10.1182/blood.v122.21.3290.3290 ◽

2013 ◽

Vol 122 (21) ◽

pp. 3290-3290

Author(s):

Qifa Liu ◽

Hui Liu ◽

Daihong Liu ◽

Yongrong Lai ◽

Jing Sun ◽

...

Keyword(s):

Overall Survival ◽

Stem Cell ◽

Research Funding ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Hematopoietic Stem ◽

Graft Versus Host ◽

Tnf Α ◽

Before And After ◽

First Complete Remission

Abstract Background Results from single institutions had shown that compared with busulfan plus cyclophosphamide (BuCy) conditioning, limiting tissue damage by myeloablative busulfan plus fludarabine (BuFlu) conditioning might decrease cytokines release, leading a lower incidence of the graft-versus-host disease (GVHD) in allogeneic hematopoietic stem cell transplantation (allo-HSCT). In our prospective, multicenter and parallel-group study, further comparison was made of the incidences and severities of GVHD following BuCy and BuFlu myeloablative conditioning regimens in patients undergoing allo-HSCT for AML in first complete remission (CR1), and analyzed plasma cytokines before and after the conditioning. Methods A total of 148 patients with AML-CR1 undergoing allo-HSCT were enrolled into BuCy (busulfan1.6mg/kg, iv q12 hours, -7 ∼ -4d; cyclophosphamide 60 mg/kg.d, -3 ∼ -2d) or BuFlu (busulfan 1.6 mg/kg, iv q12 hours, -5 ∼ -2d; fludarabine 30 mg/m2.d, -6 ∼ -2d) group between January 2007 and January 2013. For patients enrolled between January 2012 and January 2013, plasma concentrations of IL-6, IL-1β, TNF-α, CXCL-10 and IL-17A before and after conditioning were measured by Enzyme-linked immunosorbent assay (ELISA) and compared between the two groups. Regimen-related toxicity (RRT), incidences and severities of acute and chronic GVHD, and overall survival were compared between the two groups. Results Of the 148 patients enrolled in the study, the data of 142 cases were used to determine the endpoints in the intent-to-treat population (72 in BuFlu group and 76 in BuCy group). The levels of TNF-α and IL-6 were significantly higher after the conditioning (5.60±4.40 vs 8.94±5.50 and 2.19±1.24 vs 6.06±12.16 pg/ml, P <0.001 and P =0.045 ), however, there were no significant differences on these cytokines between the two groups. The levels of CXCL-10 in BuCy group was significantly higher than that in BuFlu group (P =0.012). The incidence of I-II° and III-IV° acute GVHD were 42.1% and 6.8%, and 36.1% and 5.7%, respectively, in BuCy and BuFlu group (P=0.363 and P=0.770, respectively). Chronic GVHD occurred in 29 of 69 (41.7%) and 30 of 72 (41.7%) patients, respectively, in BuCy and BuFlu group (P= 1.000). And the incidence of extensive chronic GVHD were 14.3% and 16.7%, respectively, in BuCy and BuFlu group (P= 0.670). The median follow up duration was 824 (range, 3–2345) days. The 5 year overall survival were 79.2 ± 4.4% and 78.6 ± 76.1% (P= 0.555), respectively in BuCy and BuFlu group Conclusion In this report, the incidences and severities of acute GVHD as well as chronic GVHD were similar between BuFlu and BuCy regimen in AML-CR1 patients undergoing allo-HSCT. Disclosures: Liu: National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174).: Research Funding; It was supported by 863 Program (No. 2011AA020105), National Public Health Grand Research Foundation (Grant No. 201202017): Research Funding.

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Peptides Derived from the CMV Proteome Mimic Unique Stem Cell Transplant Recipient Specific Peptides: Possible Role in Eliciting a Pro-Gvh T Cell Response

Blood ◽

10.1182/blood.v126.23.4285.4285 ◽

2015 ◽

Vol 126 (23) ◽

pp. 4285-4285

Author(s):

Charles E. Hall ◽

Max Jameson-Lee ◽

Abdelrhman Elnasseh ◽

Vishal Koparde ◽

Allison F. Scalora ◽

...

Keyword(s):

Stem Cell ◽

Amino Acid ◽

Research Funding ◽

Stem Cell Transplant ◽

Cross Reactivity ◽

Peptide Libraries ◽

Cell Transplant ◽

Graft Versus Host ◽

Sequence Identity ◽

Hla Molecules

Abstract Stem cell transplant (SCT) recipients who develop human cytomegalovirus (hCMV) reactivation are at risk of developing graft versus host disease (GVHD). This may stem from immune cross reactivity (Figure 1) towards both pathogen-derived peptides and nearly identical recipient-derived alloreactive peptide minor histocompatibility antigens from SCT donor-recipient pairs (DRP). Whole exome sequencing was performed on 9 SCT DRP, and the resulting nucleotide sequences aligned and compared, identifying all the single nucleotide polymorphisms (SNPs) present in the recipient and absent in the donor. Alloreactive peptide libraries were compiled for each DRP (9-mer peptides) and HLA binding affinity calculated for relevant HLA (Front Immunol 2014). A library representing the hCMV proteome was generated from the NCBI Protein database and directly compared to libraries of recipient specific peptide-HLA complexes from the SCT D-R pairs. BLAST Protein sequence alignment was performed to interrogate each pair's peptide-HLA complex library for matches with hCMV peptides, identifying CMV peptides with a 6/6-9/9 amino acid sequence matching to recipient peptides bound to HLA (IC50<500). The resulting matched CMV peptide libraries were completed with flanking amino acids from the original hCMV protein sequences to generate a match library of 9-mer peptides, evaluated for binding to HLA Class I using the netMHCpan algorithm 2.8 (IC50<500). Class I HLA molecules were interrogated for both alloreactive and CMV-derived peptide binding affinity in each patient. 18 HLA molecules bound both types of peptides, of which 7 HLA had more than 5 matches, i.e. human peptide(s) with 6-9 consecutive amino acid sequence identity with CMV peptide(s), where multiple matches were possible. These included: HLA-C*03:03, C*03:04, A*02:01, B*15:03, B*27:05, B*07:02 and B*15:16. Upon peptide library comparison, a median 27 (Range: 3-40) unique CMV peptides/patient matched 22 (Range: 3-30) recipient peptides from 5 matched unrelated DRP (MUD) with corresponding HLA specific binding IC50<500. Matched related DRP (MRD; n=4) comparison yielded a median 7 (Range: 1-21) unique CMV peptides/patient matching 6.5 (Range: 1-19) recipient peptides. This pattern of CMV-matched alloreactive peptides in MUD demonstrated a 3-4 fold higher degree of potential cross reactivity (Median peptides/patient: p=0.048) than MRD. MUD patients with HLA-C*03:03 & C*03:04 specificity (n=4) produced a median 20 (Range: 19-22) unique CMV peptides matching 14.5 (Range: 12-19) potentially cross-reactive recipient peptides. Cross-reactive recipient peptides were capable of matching with up to 7 different CMV peptides (Range: 1-7) and alternatively matching CMV peptides with up to 5 different recipient peptides (Range: 1-5), indicating this phenomenon could promote various strength cross-reactive immune responses (within and outside the IC50<500 range) even from a single matched CMV peptide. Tissue distribution mapping (Genotype-Tissue Expression project, GTEx) of the corresponding DRP peptide source gene expression (human mRNAs) by major tissue group indicated an overlap with known GVHD target organs from CMV-matched alloreactive recipient peptides. CMV is known to infect the vascular endothelium where cross-reactive immune cells, and potentially cross-reactive memory T cell populations, interacting with the target cells of interest would have the greatest opportunity to impact baseline inflammation and potentially trigger graft-versus-host disease (GVHD). We propose that from in silico demonstration of sequence identity between DRP polymorphic peptides with CMV proteins that immune cross-reactivity may result in those DRP and cause recipient cells to be targeted by CMV specific T cells with pro-GVH implications. Disclosures Buck: CHRB: Research Funding. Neale:CHRB: Research Funding.

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The Impact of Age in Allogeneic Stem Cell Transplantation with Thiotepa Busulfan and Fludarabine (TBF) for Myelofibrosis : A Retrospective Analysis

Blood ◽

10.1182/blood-2021-145711 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 4858-4858

Author(s):

Federica SORA ◽

Patrizia Chiusolo ◽

Sabrina Giammarco ◽

Idanna Innocenti ◽

Francesco Autore ◽

...

Keyword(s):

Stem Cell ◽

Stem Cell Transplantation ◽

Total Dose ◽

Cell Transplantation ◽

Older Patients ◽

Research Funding ◽

Acute Gvhd ◽

Conditioning Regimen ◽

Hematopoietic Stem ◽

The Impact

Abstract Allogeneic hematopoietic stem-cell transplantation (HSCT) currently remains the only curative therapy for intermediate or high risk disease.myelofibrosis (MF). We are reporting 56 patients (pts) who underwent an allogeneic HSCT in our Centre between 2016 and 2020, and assessed factors predictive of outcome. The median age was 59 years (36-72). Most patients (72%) were JAK2+ and had int2-high DIPSS (92%). The conditioning regimen consisted of thiotepa, busulfan , fludarabine (TBF). All pts received thiotepa 10 mg/kg and fludarabine 150 mg/m^2. The dose of busulfan was adjusted considering the age and the comorbidity score. One pt received 3 days of busulfan (total dose 9.6 mg/kg); 47 received 2 days (total dose 6.4 mg/kg) and 8 received one day of busulfan iv (3.2 mg/kg). Donor was an identical sibling in 13 pt, haploidentical in 18, matched unrelated donor (UD) in 18 and a mismatchedUD in 7. Thus we had 31 HLA matched and 25 HLA mismatched grafts. Fortytwo patients received post-transplant cyclophosphamide (PTCy)-based GVHD (Graft versus host disease ) prophylaxis with cyclosporine and mycophenolate mofetil , and 14 patients received a standard GvHD prophylaxis (CSA+MTX+ATG). The 2 year survival (OS) was 73 % and disease free survival (DFS) was 66 % and the cumulative incidence (CI) of TRM was 23% and of relapse 11%. The incidence of acute GvHD grade II-IV was 22% in HLA matched and 50% in HLA mismatched pts (p=0.022), grade III-IV was 6% and 25% respectively (p=0.042) . The incidence of moderate-severe chronic GvHD was 25% in HLA matched and 36% in HLA mismatched grafts (p=0.36). HLA had a major impact on survival : 85% vs 49% survival for matched vs mismatched patients (p=0.01). Patients age >60 years had a major impact on outcome, with a 2 year survival of 51% vs 88% in patients over (n=24) or under 60 years of age (n=32) (p=0.007; the DFS was 46 % and 80% respectively and the CI of TRM was 42% vs 9% (p=0.003). As to the total dose of busulfan, we found 26% TRM in patients receiving busulfan for 2 days (total doe 6.4 mg/kg) (n=47) and 0% in older patients receiving 1 day only (total dose 3.2 mg/kg) (n=8) ; relapse rate was 10% and 20% respectively. In multivariate cox analysis including age, spleen size ,DIPSS score, number of transfusion received and donor type, only HLA matching influenced the incidence of acute GvHD; transfusion burden and age plays a role in NRM and OS; DIPSS predicts relapse . In conclusion: older patients with MF have a high NRM and need to be prepared with a milder conditioning regimen. Disclosures Laurenti: Janssen: Consultancy, Honoraria; AstraZeneca: Consultancy, Honoraria; AbbVie: Consultancy, Honoraria, Research Funding; Roche: Honoraria, Research Funding; Gilead: Honoraria; BeiGene: Honoraria. Sica: Pfizer: Honoraria.

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Dose Study of Antithymocyteglobulin in Haploidentical Allogeneic Stem Cell Transplantation

Blood ◽

10.1182/blood.v124.21.1164.1164 ◽

2014 ◽

Vol 124 (21) ◽

pp. 1164-1164

Author(s):

Qifa Liu ◽

Ren Lin ◽

Zhiping Fan ◽

Qianli Jiang ◽

Min Dai ◽

...

Keyword(s):

Stem Cell ◽

Cumulative Incidence ◽

Research Funding ◽

Acute Gvhd ◽

Guangdong Province ◽

Unrelated Donor ◽

Total Dosage ◽

Guangzhou City ◽

To Receive ◽

Neutrophil Engraftment

Abstract Backgroud:Graft-versus-host diseases (GVHD) remains a major complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT), especially in recipients with HLA-mismatched or unrelated donors. Antithymocyteglobulin (ATG) has been widely used to prevent acute GVHD (aGVHD) in haploidentical and unrelated donor transplantation The use of ATG may increase the risk of opportunistic infections and primary disease relapse, which is associated with the dose of ATG. Till now, the optimal dose of ATG is not known for haploidentical and unrelated donor transplantation. Here, we compared the outcome of patients undergoing haploidentical HSCT who were treated with two different doses of ATG. Methods: Between January 2013 and June 2014, 40 consecutive patients with hematological malignancies undergoing haploidentical HSCT were enrolled in this prospective study. Of the 40 patiens, 19 received allo-HSCT from sibling donors, 18 lineal relatives donors (i.e. father, mother or child) and 3 collateral relatives donors (i.e. uncle or aunt). Five patients received 1 locus HLA-A, -B, -DRB1, -C, -DQ-mismatched donor transplantation, 6 received 2 locus mismatched, 9 received 3 locus mismatched, 6 received 4 locus mismatched and 14 received 5 locus mismatched transplantation. All the patients received GVHD prophylaxis including cyclosporine A, short-term methotrexate (on day +1, +3, and +6) and mycophenolate mofetil (0.5 g twice a day on day 0 to +28) as well as ATG ( a total dosage of 7.5mg/kg or 10mg/kg according to randomization). Epstein-Barr virus (EBV) -DNA and cytomegalovirus (CMV) -DNA levels of blood were monitored weekly for the first 3 months after transplantation, every two weeks during the 4 to 9 month after HSCT, and then once a month during 10 to 12 month. The primary endpoint was the cumulative incidence of aGVHD within day +100 after allo-HSCT. Results:Nineteen of the 40 patients were randomized to receive a total dosage of 7.5 mg/kg ATG and 21 were randomized to receive 10 mg/kg ATG. One patient was withdrawn from transplantation because of infection during conditioning and did not receive ATG. Of the 39 evaluable patients, 38 achieved engraftment except for one died of infection on day +32. The median time to neutrophil engraftment was 13 days (range, 9 to 19 days), and the median time to platelet engraftment was 14 days (range, 10 to 73 days). Patients with 7.5 mg/kg ATG achieved neutrophil engraftment earlier than those with 10 mg/kg (P=0.011). Time to platelet engraftment was comparable in the 2 arms with different dosage of ATG (P=0.063). The cumulative incidence of aGVHD grades II to IV and III to IV within day 100 post-transplantation was 26.5±7.2% and 10.6±5.0%, respectively. Acute GVHD grade II to IV developed in 35.5±11.8% of the patients with 7.5 mg/kg ATG and 19.3±8.7% of those with 10 mg/kg ATG (P=0.273). The incidence of aGVHD grade III to IV within day 100 were 11.9±7.9% in 7.5 mg/kg arm and 9.8±6.6% in 10 mg/kg arm (P=0.831). The cumulative incidence of chronic GVHD (cGVHD) 50.0%±25.0% and 59.6%±21.1% in the patients with 7.5 mg/kg and 10mg/kg ATG, respectively (P=0.819). The 1-year cumulative incidence of CMV reactivation were similar in the two arms (7.5 mg/kg arm : 82.8±10.5% vs 10 mg/kg arm: 66.7±10.3%, P=0.600). The incidence of EBV reactivation were 49.6±12.5% in 7.5 mg/kg arm and 67.2±12.7% in 10 mg/kg arm (P=0.729).Six patients relapsed, including 3 receiving 7.5 mg/kg ATG and 3 receiving 10 mg/kg ATG. The median follow up was 247 days (range, 32 to 569 days). The 1-year cumulative overall survival were 62.8±12.6% in 7.5 mg/kg arm and 68.2±10.9% in 10 mg/kg arm (P=0.536). The 1-year cumulative non-relapse mortality (NRM) rates were 23.4±10.3% and 23.3±10.5% in 7.5 mg/kg and 10 mg/kg arm, respectively (P=0.609). Conclusion:This trial suggests that 7.5 mg/kg ATG might have similar efficacy in preventing aGVHD after haploidentical HSCT compared with 10 mg/kg.Whether patients with 7.5 mg/kg ATG have lower risk for viral infections than those with 10 mg/kg needs further studies. Disclosures Liu: National Natural Science Foundation of China (81270647, 81300445, 81200388): Research Funding; National High Technology Research and Development Program of China (863 Program) (2011AA020105): Research Funding; National Public Health Grand Research Foundation (201202017): Research Funding; Natural Science Foundation of Guangdong Province (S2012010009299): Research Funding; the project of health collaborative innovation of Guangzhou city (201400000003-4, 201400000003-1): Research Funding; the Technology Plan of Guangdong Province of China (2012B031800403): Research Funding; the project of the Zhujiang Science & Technology Star of Guangzhou city (2013027): Research Funding.

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The Effect of JAK 1/2 Inhibitors on Outcomes of Allogeneic Stem Cell Transplantation for Patients with Myelofibrosis

Blood ◽

10.1182/blood-2018-99-112769 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 5784-5784

Author(s):

Guido Lancman ◽

Kathleen Miller ◽

Shuli Li ◽

Vincent T. Ho ◽

Amir T. Fathi ◽

...

Keyword(s):

Stem Cell ◽

Stem Cell Transplantation ◽

Board Of Directors ◽

Cell Transplantation ◽

Allogeneic Stem Cell Transplantation ◽

Research Funding ◽

Acute Gvhd ◽

Advisory Committees ◽

Free Survival ◽

Donor Type

Abstract Introduction: Ruxolitinib was the first JAK 1/2 inhibitor (JAKi) approved for myelofibrosis (MF), with several other JAKi in development. Ruxolitinib was approved on the basis of reducing splenomegaly and improving constitutional symptoms, but its effect on subsequent allogeneic stem cell transplantation (SCT) is not well understood. Retrospective studies to date have reported mixed outcomes after SCT for MF patients with previous exposure to JAKi. In this multicenter retrospective study, we report on outcomes of patients with MF treated with SCT at our institutions. Methods: We analyzed outcomes for 184 consecutive patients at three institutions who underwent SCT for primary or secondary MF. Primary outcomes included overall survival (OS), progression free survival (PFS), and graft-versus-host-disease (GVHD)-free and relapse-free survival (GRFS), all measured from the time of SCT. Cox proportional hazard regressions were fit to estimate the association between the use of JAK 1/2 inhibitors prior to SCT and OS, PFS, and GRFS, adjusting for donor type and DIPSS-plus status. p<0.05 was considered statistically significant. Results: 72 patients received a JAKi prior to SCT, while 112 did not. Patients in these two groups were well-matched with respect to age, sex, DIPSS plus score, conditioning, and donor type (Table 1). Median follow-up was 31.2 months (range: 0.8-146.3 months). In univariate analysis, there was no difference in OS (JAKi: 4-yr OS 56.7% [95% CI 40.9-69.8%] vs. no JAKi: 43.6% [95% CI 32.9-53.9%], p=0.49), PFS (JAKi: 4 yr PFS 54.1% [95% CI 40.8-65.7%] vs. no JAKi: 43.9% [95% CI 33.4-53.9%], p=0.77), or GRFS (JAKi: 8-month GRFS 56.6% [95% CI 44.1-67.4%] vs. no JAKi: 50.4% [95% CI 40.4-59.5%], p=0.62) in the overall population; there was similarly no difference when comparing only intermediate-risk or only high-risk patients. In multivariate analysis, there was no difference in these outcomes for patients based on previous JAKi exposure when accounting for DIPSS plus score and donor type (related vs unrelated). Rates of acute GVHD were similar between the two groups (JAKi: 53.5% vs. no JAKi: 55.0%, p=0.88), including grade 3 or 4 acute GVHD (JAKi: 16.9% vs no JAKi: 19.8%, p=0.70). Conclusions: Our data suggest that there is no statistically significant difference in OS, PFS, GRFS, or rates of acute GVHD after SCT for MF patients based on previous JAKi treatment. This was true overall and after adjusting for DIPSS plus risk score or donor type. Given the retrospective design of our study, we were not able to assess prior response to JAKi or splenomegaly at SCT, which may influence outcomes. Given mixed results in the literature to date, we eagerly await the results of ongoing phase 2 trials of JAKi prior to SCT for MF. Disclosures Ho: Jazz Pharmaceuticals: Consultancy. Fathi:Astellas: Honoraria; Jazz: Honoraria; Boston Biomedical: Consultancy, Honoraria; Takeda: Consultancy, Honoraria; Agios: Honoraria, Research Funding; Celgene: Consultancy, Honoraria, Research Funding; Seattle Genetics: Consultancy, Honoraria. Chen:Takeda Pharmaceuticals: Consultancy; Incyte: Consultancy, Membership on an entity's Board of Directors or advisory committees; Magenta Therapeutics: Consultancy; REGiMMUNE: Consultancy. Hoffman:Formation Biologics: Research Funding; Incyte: Research Funding; Janssen: Research Funding; Merus: Research Funding; Summer Road: Research Funding. Mascarenhas:Novartis: Research Funding; Merck: Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees; Incyte: Membership on an entity's Board of Directors or advisory committees, Research Funding; Promedior: Research Funding; Janssen: Research Funding; Roche: Research Funding; CTI Biopharma: Membership on an entity's Board of Directors or advisory committees, Research Funding.

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Prior Use of Mogamulizumab to Allogenic Hematopoietic Stem Cell Transplantation Induces Severe Acute Graft-Versus-Host Disease

Blood ◽

10.1182/blood.v126.23.1940.1940 ◽

2015 ◽

Vol 126 (23) ◽

pp. 1940-1940 ◽

Cited By ~ 1

Author(s):

Takeshi Sugio ◽

Koji Kato ◽

Takatoshi Aoki ◽

Takanori Ota ◽

Noriyuki Saito ◽

...

Keyword(s):

Bone Marrow ◽

T Cells ◽

Stem Cell ◽

T Cell ◽

Regulatory T Cells ◽

Research Funding ◽

Acute Gvhd ◽

Cell Lymphoma ◽

Hematopoietic Stem ◽

Graft Versus Host

Abstract [Introduction] Adult T-cell leukemia/lymphoma (ATL) is an aggressive peripheral T-cell lymphoma (PTCL) with a dismal prognosis. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the only curative treatment in ATL patients. Mogamulizumab, a humanized anti-CC chemokine receptor 4 (CCR4) monoclonal antibody, is a novel immunotherapeutic agent, effective in treating patients with PTCL such as ATL, PTCL-not specified, and cutaneous T-cell lymphoma. However, in allo-HSCT setting, we should be careful to use mogamulizumab because CCR4 is expressed in regulatory T cells: The mogamulizumab treatment may accelerate GVHD by eradicating regulatory T cells in allo-HSCT patients. Here, we retrospectively analyzed the effect of mogamulizumab on GVHD development in ATL patients treated with mogamulizumab prior to allo-HSCT. [Patients and Methods] Data from the Fukuoka Bone Marrow Transplantation Group were retrospectively analyzed after the approval of mogamulizumab use in Japan. [Results] A total of 24 patients with ATL received mogamulizumab prior to allo-HSCT between April 2012 and April 2015 in our group. The median age at allo-HSCT was 58.5 years (range, 32-72). The median intervals from the last administration of mogamulizumab to allo-HSCT were 25 days (range, 9-126). The median total dose of mogamulizumab was 3 mg/kg (range, 1-8 mg/kg). After treatment with mogamulizumab, 18 patients (75%) had achieved in remission (CR in 4 patients and PR in 14) at allo-HSCT. Ten patients received unrelated bone marrow, 5 received related peripheral blood, and 9 received cord blood as stem cell sources. Eleven patients were treated with full-intensity conditioning and 13 received reduced-intensity conditioning. Graft-versus-host disease (GVHD) prophylaxis consisted of calcineurin inhibitors (cyclosporine or tacrolimus) with short-term methotrexate in 14 patients and mycophenolate mofetil in 9. The cumulative incidence (CI) of acute GVHD at 100 days was 66.6% in grade 2-4 and 33.3% in grade 3-4. The involved organs of acute GVHD were skin in 14 patients, gut in 10, and liver in 4. Among 14 patients who developed grade 2-4 acute GVHD, 5 had severe fluid retention such as pleural effusion or ascites associated with GVHD. Chronic GVHD was observed in 6 patients, and 5 of them were extensive disease. The CI of transplant-related mortality (TRM) and relapse at 1-year were 53.2% (95%CI, 29.3-72.3%) and 29.6% (95%CI, 12.6-48.9%), respectively. The leading cause of death was GVHD (n = 7). The 1-year overall survival and progression-free survival were 19.2% (95%CI, 5.7-38.8%) and 17.2% (95%CI, 4.9-35.7%), respectively. [Discussion] Use of mogamulizumab prior to transplantation in allo-HSCT patients has a merit to decrease the burden of ATL cells. However, it was associated with an increase of TRM due to severe GVHD. Although most of ATL patients achieved better disease status at allo-HSCT through mogamulizumab and the survival rate was expected to be 50% based on the previous data, the survival in the present study was ~20%. These data suggest that mogamulizumab administered before transplantation may have retained until an early phase of post-transplantation, and the donor or host-derived regulatory T cells might be eliminated, allowing the GVHD T-cell clone to expand. Since mogalizumab is a potent anti-ATL agent, we need to develop new treatment protocols integrating mogalizumab at a suitable dose or administration timing, to minimize the unwanted GVHD development in future studies. Disclosures Akashi: Asahi Kasei: Research Funding, Speakers Bureau; Shionogi: Research Funding, Speakers Bureau; Astellas: Research Funding, Speakers Bureau; Celgene: Research Funding, Speakers Bureau; Chugai: Research Funding, Speakers Bureau; Bristol-Myers Squibb: Research Funding, Speakers Bureau; Novartis Pharma K.K.: Consultancy, Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Consultancy, Research Funding, Speakers Bureau.

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Evaluation of Graft Versus Host Disease and Relapse Free Survival As Novel Endpoint in Allogeneic Hematopoietic Stem Cell Transplantation: A Retrospective Joint Naples-Paris Study

Blood ◽

10.1182/blood.v128.22.2285.2285 ◽

2016 ◽

Vol 128 (22) ◽

pp. 2285-2285

Author(s):

Simona Pagliuca ◽

Antonio M Risitano ◽

Sylvie Chevret ◽

Flore Sicre de Fontbrune ◽

Alienor Xhaard ◽

...

Keyword(s):

Stem Cell ◽

Cumulative Incidence ◽

Research Funding ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Relapse Free Survival ◽

Hematopoietic Stem ◽

Free Survival ◽

Graft Versus Host

Abstract The cure of hematologic disorders by allogeneic hematopoietic stem cell transplantation (HSCT) is often associated with major complications resulting in poor outcome, including acute and chronic graft-versus-host disease (GVHD), relapse and death. Classical endpoints such as overall survival (OS), desease free survival (DFS) and non relapse-mortality (NRM) had become more and more unsuitable for transplant research because of their inability to a dynamic mesure of transplant-associated comorbidity. For this reason several composite endpoints taking into account also GVHD-associated comorbidity were proposed in the last years. GVHD free/relapse free survival (GRFS), proposed by Holtan et al (Blood 2015), includes grades 3-4 acute GVHD, systemic therapy requiring chronic GVHD, primary disease relapse , or death for any cause considered as events. This endpoint seems to completely characterize the survival without mortality or ongoing morbidity. With the intent to analyse the outcomes of our transplanted cohort, we retrospectively analysed GRFS of 959 consecutive patients receiving HSCT at Federico II University in Naples (n=119) and Saint-Louis Hospital (n=840) in Paris between 2007 and 2014, identifying prognostic factors associated with a better outcome and estimating the incidences of all components of this endpoint: rates of acute and chronic GVHD, disease relapse and death. Patient, disease and transplant characteristics are listed in table 1. Median duration of follow-up after HSCT was 22.1 months (IQR: 5.6-51 months). Cumulative incidence at day 100 of grade II-IV acute GVHD and grade III-IV were 42% and 16%, respectively. Cumulative incidence of chronic GVHD requiring systemic treatment at 1 and 5 years was 23% and 33%, respectively, diagnosed according to NIH criteria [14% of patients had score 1 (mild), 58% score 2 (moderate) and 27% score 3 (severe)cGVHD]. Cumulative incidence of relapse (considering all malignant and non-malignant diseases) was 26.7% (N=219) at 5 years. Overall survival for the whole population was 57% (95%CI, 53.3-60.8) at 5 years and Disease free survival (DFS) and non-relapse mortality (NRM) were respectively 50% (95%CI, 46.6-53.8) and 23% at 5 years. GRFS was 25% (95%CI, 21.8-28.5) at 5 years. Factors identified as influencing GRFS based on univariate analyses were age higher than 45 years (HR=1.64, 95%CI, 1.40-1.92), bone marrow (BM) as stem cell source (HR=0.40, 95%CI, 0.32-0.50); reduced intensity conditioning (RIC) (HR=0.63, 95%CI, 0.53-0.74); disease type [non-malignant disorders: HR=0.24, 95%CI, 0.17-0.33; myelodysplastic and myeloproliferative syndromes (MPN/CML/MDS): HR=1.34, 95%CI, 1.10-1.63; whereas other diagnosis did not influence GRFS] and than unrelated donor (matched: HR=1.71, 95%CI, 1.41-2.07;mismatch:HR=1.81, 95%CI, 1.48-2.23). Based on a multivariable Cox model, only diagnoses (non-malignancies, HR=0.27, 95%CI, 0.19-0.38 and MPN/CML/MDS, HR= 1.35, 95%CI, 1.11-1.65), and HLA unrelated graft (matched, HR=1.42, 95%CI, 1.17-1.73 and mismatched, HR=1.55, 95%CI, 1.26-1.92) remained associated with the outcome (Figure 1 and 2). GRFS could represent the ideal endpoint following HSCT. It differs significantly based upon type of disease and donor type, essentially. This composite indicator yields more information regarding complications of HSCT than the simpler measurement of OS or DFS. Its use willbetter compare these clinically important outcomes that accompany disparate HSCT techniques. All examined prognostic factors could enhance our ability to optimally judge the risk and the probability of true recovery after allogeneic HSCT. Our data support the use of this composite endpoint to describe HSCT outcome, and also pave the way for the investigation of novel endpoints, which may also track the dynamic changes of post-transplant events in the long-term. These retrospective data represent the background to investigate the impact of novel strategies of HSCT aiming to improve the outcome of HSCT, as detectable, by using more sensitive endpoints, tracking clinical events associated with detrimental long-term outcome. Figure 1 Figure 1. Figure 2 Figure 2. Disclosures Risitano: Alexion Pharmaceuticals: Other: lecture fees, Research Funding; Novartis: Research Funding; Alnylam: Research Funding; Rapharma: Research Funding. Peffault de Latour:Pfizer: Consultancy, Honoraria, Research Funding; Amgen: Research Funding; Novartis: Consultancy, Honoraria, Research Funding; Alexion: Consultancy, Honoraria, Research Funding.

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Decentralized Manufacture of TCR-Alpha/Beta and CD19 Depleted Haploidentical Stem Cell Grafts for Children within a Multicenter Phase I/II Clinical Trial

Blood ◽

10.1182/blood.v128.22.2172.2172 ◽

2016 ◽

Vol 128 (22) ◽

pp. 2172-2172

Author(s):

Michael Schumm ◽

Matthias K. Eyrich ◽

Markus Wiesneth ◽

Halvard Bönig ◽

Peter J. Lang ◽

...

Keyword(s):

Stem Cell ◽

B Cells ◽

Research Funding ◽

Pediatric Patients ◽

Acute Gvhd ◽

Round Robin ◽

Cell Numbers ◽

Cd34 Cells ◽

Alpha Beta

Abstract Several single center experiences have shown favorable outcomes using in-vitro depletion of T cell receptor (TCR)-alpha/beta cells and B cells. For the first time we show the manufacturing results of stem cell grafts from haploidentical family donors for 30 pediatric patients with various hematological and non-hematological malignancies and non-malignant diseases within a prospective, multi-center phase I/II clinical trial utilizing the CliniMACS plus System (Miltenyi Biotec, Germany) in combination with a reduced conditioning (www.clinicaltrialsregister.org; 2011-005562-38). The in-vitro T cell depletions of the grafts were performed by four different laboratories under fixed conditions. The grafts were sent to six treatment centers. Methods: Donors received G-CSF for mobilization of stem cells according to local hospital routine followed by leukaphereses that were depleted from TCRab+ and CD19+ cells according to manufacturer's instructions (CliniMACS plus System, Miltenyi Biotec) and as approved by local authorities. For quality purposes the sponsor performed regularly round robin tests to ensure provision of comparable results regarding the residual number of TCRab+ cells in the grafts. The transplants should be composed of a maximum of 7.5 x 108 nucleated cells/mL with targeted ≥ 4 x 106 CD34+ cells/kg, ≤ 25 x 103 TCRab+ cells/kg, ≤ 1 x 105 CD20+ cells/kg and a CD34+ cell viability of ≥ 95%. In case the targeted value for CD34+ cells/kg was not reached, the number of TCRab+ cells/kg was allowed to exceed by up to the four-fold until 4 x 106 CD34+ cells/kg per transplant have been isolated. One transplant could consist of up to three single aphereses depleted of TCRab+ and CD19+ cells. These products were to be infused directly after separation or following cryopreservation. Pooling of aphereses from a single donor was not allowed. Results: 30 pediatric patients, median age 7 years (range, 1 - 17 years) received a total of 43 TCRab/CD19 depleted haploidentical stem cell products. 17 patients got one infusion, 11 two and one patient three infusions. The stem cell products contained in median 9.46 x 106/kg (range, 1.35 - 54.9) CD34+ cells, 8.4 x 103/kg (range, 0.62 - 40.6) TCRab+ cells and 0.32 x 105/kg (range, 0.037 - 1.7) CD20+ cells. Log depletion for TCRab+ cells and B cells was 4.75 (range, 1.2 - 5.33) and 3.43 (range, 0 - 3.93), respectively. In addition significant numbers of NK and TCRgd+ cells/kg were preserved: 3.9 x 107 (median; range, 0.11 - 18.2), 0.67 x 107 (median; range, 0.05 - 4.0), respectively, summing up to a median number of 6.99 x 106(range, 0.42 - 39.7) total CD3+ cells/kg. Viability of CD34+ cells was 97.9% (median, range 91.5 - 100). All patients received transplants (consisting of up to three single consecutive products) with the targeted CD34+ cell dose - median 14.6 x 106 cells/kg (range, 4 - 54.9) and with less than the maximal allowed number of 1x105 TCRab+ cells/kg. Three single products exceeded targeted TCRab cell numbers but remained within the defined limit of the transplant of 1 x 105 cells/kg in order to meet the specification of ≥ 4 x 106 CD34+ cells/kg. In six single products the B cell numbers were above the specified targeted limit (max. 1.7 x 105/kg). Four products had a viability of CD34+ cells between 91.5 and 95%. Of 30 treated patients no patient experienced acute GVHD°III-IV. Only one patient had acute GVHD°II. The transplant of this patient fulfilled the targeted specifications for total CD34+, TCRab+ and CD20+ cells/kg. Round robin tests were performed prior to study start and during the enrollment period. The identified issues were addressed prior to study start and subsequent tests revealed a uniform performance of the manufacturing centers. Conclusion: 43 stem cell products were manufactured and released by 4 manufacturing laboratories for 30 pediatric patients in 6 hospitals within a clinical study investigating TCR alpha/beta and CD19 depleted haploidentical stem cell transplantation after reduced intensity conditioning. A highly effective depletion of TCRab + cells and B cells with comparable results was shown in all laboratories as controlled by frequent evaluation in round robin tests. Limited exceedances of the targeted release criteria of TCRa/b+ cells were acceptable to the physicians and had no clinical impact. Disclosures Bönig: Miltenyi Biotec: Consultancy, Honoraria, Research Funding. Bader:Riemser: Research Funding; Neovii Biotech: Research Funding; Servier: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Medac: Consultancy, Research Funding. Aktas:Miltenyi Biotec: Employment. Dresing:Miltenyi Biotec: Employment. Karitzky:Miltenyi Biotec: Employment. Holtkamp:Miltenyi Biotec: Employment. Handgretinger:Miltenyi Biotec: Patents & Royalties: Co-Patentholder of TcRalpha/beta depletion technology.

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Long-Term Survival after Transplantation in Patients with Chronic Myeloid Leukemia from HLA-Compatible Unrelated Donors: May the Stem Cell Source Influence the Outcome?

Blood ◽

10.1182/blood-2018-99-118615 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 5775-5775

Author(s):

Michael Koldehoff ◽

Ahmet H. Elmaagacli ◽

Hellmut Dietmar Ottinger ◽

Ulrich Dührsen ◽

Dietrich W. Beelen

Keyword(s):

Stem Cell ◽

Myeloid Leukemia ◽

Research Funding ◽

Acute Gvhd ◽

Peripheral Blood Stem Cell ◽

Chronic Gvhd ◽

Disease Stage ◽

Unrelated Donor ◽

Advanced Stages

Abstract Introduction Allogeneic hematopoietic stem cell transplantation (alloSCT) is curative therapy for chronic myeloid leukemia (CML), but its long-term outcomes regarding graft sources are not well described. Results Here the outcomes of CML after alloSCT with bone marrow (BM, n=134) were compared to those of peripheral blood stem cell (PBSC, n=172) in the HLA-compatible unrelated donor setting. Patients were transplanted in 1st CP (Bone marrow transplantation (BMT), n=100, and Peripheral blood stem cell transplantation (PBSCT), n=103), in >1st CP (BMT, n=24, and PBSCT, n=52) and in blast crises (BMT, n=10, and PBSCT, n=17). Median follow-up were 54 months after BMT and 106 months after PBSCT. No significant differences were found in the incidence of acute and chronic graft-versus-host disease (GvHD) between both study-groups. The 5-year estimated probability of hematological relapse was 11% for patients in 1st CP CML and 49% for patients in advanced disease after BMT (p<0.001) and 18% for patients in 1st CP and 22% for patients in advanced disease after PBSCT (p= n.s.). The estimated probability for 10-year overall survival (OS) for patients in 1st CP and patients in advanced stages of CML were 58% and 20% after BMT and 59% and 52% after PBSCT, respectively (not significant for 1st CP and p<0.0001 for advanced stages). In the multivariate analysis patient age (age > 40 years), disease stage, acute GvHD, chronic GvHD and immunprophylaxis with use of ATG influenced OS significantly. For leukemia-free survival, the following risk factors were significantly in the multivariate analysis, graft source, patient age, gender constellation, disease stage, acute GvHD and chronic GvHD. Conclusions This large trial show significant difference between transplant recipients who received PBSC and those who received BM from unrelated donor. These finding may influence the selection of a graft source for alloSCT from unrelated donor. Disclosures Dührsen: Amgen: Research Funding; Gilead: Consultancy, Honoraria; Janssen: Honoraria; AbbVie: Consultancy, Honoraria; Roche: Honoraria, Research Funding; Celgene: Honoraria, Research Funding. Beelen:Medac: Consultancy, Other: Travel Support.

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