MESENCHYMAL STROMAL CELLS GENERATED FROM POOLED MONONUCLEAR CELLS OF MULTIPLE BONE MARROW DONORS AS A RESCUE THERAPY FOR SEVERE STEROID-REFRACTORY GRAFT VERSUS HOST DISEASE: A MULTICENTER SURVEY
(Abstract release date: 05/19/16)
EHA Library. Kuçi Z. 06/11/16; 135270; S514
Dr. Zyrafete Kuçi
Contributions
Contributions
Abstract
Abstract: S514
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:45 - 17:00
Location: Hall C14
Background
Mesenchymal stromal cells (MSCs) represent a heterogeneous cell population concerning their proliferative, differentiation and allosuppressive potential.
Aims
To circumvent donor-to-donor heterogeneity which may lead to inconsistent results after treatment of acute graft-versus-host disease (aGvHD) with mesenchymal stromal cells (MSCs) generated from a single or several individual donors, we developed a novel approach by generating MSCs from pooled bone marrow mononuclear cells (BM-MNCs) of eight healthy “third-party” donors.
Methods
BM-MNCs of 8 healthy “third-party” donors were isolated and frozen separately in cryobags. After thawing, BM-MNCs of 8 donors were pooled and cultured for 14 days in DMEM+5% platelet lysate (PL). Generated MSCs were frozen in 209 cryovials ȧ 1.5x10e6 MSCs, representing the MSC-bank. Several vials were expanded for 12-14 days in DMEM+10% PL till the end of passage 2 and frozen as MSC-end products (MEPs). These products were evaluated for proliferative, differentiation and allosuppressive potential as well as genomic stability. Eighty-one MEPs generated from aliquots of the MSC bank were administered on a compassionate-use basis after approval of the regulatory authorities to 26 patients with steroid-refractory acute GvHD at a target dose of 1–2×106 MSCs/kg BW in 7 transplantation centers.
Results
MEPs exhibited typical MSC phenotype, trilineage differentiation potential and replicative senescence after 13 passages, as determined by increased expression of senescence-related markers such as p21 (CDKN1A) and p16 (CDKN2A). Chromosomal analysis of MEPs demonstrated a normal karyotype, whereas FISH analysis demonstrated a normal diploid pattern, indicating their genomic stability. Importantly, MEPs exerted a significantly higher allosuppressive potential than the mean allosuppressive potential of MSCs generated from the same donors individually. Administration of 81 MEPs to 26 patients with severe steroid-resistant aGvHD in 7 stem cell transplant centers who were refractory to many lines of treatment, induced a 77% overall response at the primary endpoint (day 28). Remarkably, although the cohort of patients was highly challenging (92% grade III/IV and only 8% grade II GvHD), after treatment with MEPs the overall survival rate at 2 years follow-up was 71±11% for the entire patient cohort, compared to 51.4±9.0% in GvHD clinical studies, in which MSCs were derived from single donors.
Conclusion
To our knowledge this is the first serum-free MSC bank generated from pooled BM-MNCs of multiple donors as a source for bulk production of clinical-grade MSCs with a predictable potency. Importantly, clinical data presented in this study demonstrated the in vivo safety and efficacy of MEPs, which may provide a novel therapeutic tool for the effective treatment of severe aGVHD.
Session topic: Gene therapy, cellular immunotherapy and vaccination
Keyword(s): Graft-versus-host disease (GVHD), Mesenchymal stem cell
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:45 - 17:00
Location: Hall C14
Background
Mesenchymal stromal cells (MSCs) represent a heterogeneous cell population concerning their proliferative, differentiation and allosuppressive potential.
Aims
To circumvent donor-to-donor heterogeneity which may lead to inconsistent results after treatment of acute graft-versus-host disease (aGvHD) with mesenchymal stromal cells (MSCs) generated from a single or several individual donors, we developed a novel approach by generating MSCs from pooled bone marrow mononuclear cells (BM-MNCs) of eight healthy “third-party” donors.
Methods
BM-MNCs of 8 healthy “third-party” donors were isolated and frozen separately in cryobags. After thawing, BM-MNCs of 8 donors were pooled and cultured for 14 days in DMEM+5% platelet lysate (PL). Generated MSCs were frozen in 209 cryovials ȧ 1.5x10e6 MSCs, representing the MSC-bank. Several vials were expanded for 12-14 days in DMEM+10% PL till the end of passage 2 and frozen as MSC-end products (MEPs). These products were evaluated for proliferative, differentiation and allosuppressive potential as well as genomic stability. Eighty-one MEPs generated from aliquots of the MSC bank were administered on a compassionate-use basis after approval of the regulatory authorities to 26 patients with steroid-refractory acute GvHD at a target dose of 1–2×106 MSCs/kg BW in 7 transplantation centers.
Results
MEPs exhibited typical MSC phenotype, trilineage differentiation potential and replicative senescence after 13 passages, as determined by increased expression of senescence-related markers such as p21 (CDKN1A) and p16 (CDKN2A). Chromosomal analysis of MEPs demonstrated a normal karyotype, whereas FISH analysis demonstrated a normal diploid pattern, indicating their genomic stability. Importantly, MEPs exerted a significantly higher allosuppressive potential than the mean allosuppressive potential of MSCs generated from the same donors individually. Administration of 81 MEPs to 26 patients with severe steroid-resistant aGvHD in 7 stem cell transplant centers who were refractory to many lines of treatment, induced a 77% overall response at the primary endpoint (day 28). Remarkably, although the cohort of patients was highly challenging (92% grade III/IV and only 8% grade II GvHD), after treatment with MEPs the overall survival rate at 2 years follow-up was 71±11% for the entire patient cohort, compared to 51.4±9.0% in GvHD clinical studies, in which MSCs were derived from single donors.
Conclusion
To our knowledge this is the first serum-free MSC bank generated from pooled BM-MNCs of multiple donors as a source for bulk production of clinical-grade MSCs with a predictable potency. Importantly, clinical data presented in this study demonstrated the in vivo safety and efficacy of MEPs, which may provide a novel therapeutic tool for the effective treatment of severe aGVHD.
Session topic: Gene therapy, cellular immunotherapy and vaccination
Keyword(s): Graft-versus-host disease (GVHD), Mesenchymal stem cell
Abstract: S514
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:45 - 17:00
Location: Hall C14
Background
Mesenchymal stromal cells (MSCs) represent a heterogeneous cell population concerning their proliferative, differentiation and allosuppressive potential.
Aims
To circumvent donor-to-donor heterogeneity which may lead to inconsistent results after treatment of acute graft-versus-host disease (aGvHD) with mesenchymal stromal cells (MSCs) generated from a single or several individual donors, we developed a novel approach by generating MSCs from pooled bone marrow mononuclear cells (BM-MNCs) of eight healthy “third-party” donors.
Methods
BM-MNCs of 8 healthy “third-party” donors were isolated and frozen separately in cryobags. After thawing, BM-MNCs of 8 donors were pooled and cultured for 14 days in DMEM+5% platelet lysate (PL). Generated MSCs were frozen in 209 cryovials ȧ 1.5x10e6 MSCs, representing the MSC-bank. Several vials were expanded for 12-14 days in DMEM+10% PL till the end of passage 2 and frozen as MSC-end products (MEPs). These products were evaluated for proliferative, differentiation and allosuppressive potential as well as genomic stability. Eighty-one MEPs generated from aliquots of the MSC bank were administered on a compassionate-use basis after approval of the regulatory authorities to 26 patients with steroid-refractory acute GvHD at a target dose of 1–2×106 MSCs/kg BW in 7 transplantation centers.
Results
MEPs exhibited typical MSC phenotype, trilineage differentiation potential and replicative senescence after 13 passages, as determined by increased expression of senescence-related markers such as p21 (CDKN1A) and p16 (CDKN2A). Chromosomal analysis of MEPs demonstrated a normal karyotype, whereas FISH analysis demonstrated a normal diploid pattern, indicating their genomic stability. Importantly, MEPs exerted a significantly higher allosuppressive potential than the mean allosuppressive potential of MSCs generated from the same donors individually. Administration of 81 MEPs to 26 patients with severe steroid-resistant aGvHD in 7 stem cell transplant centers who were refractory to many lines of treatment, induced a 77% overall response at the primary endpoint (day 28). Remarkably, although the cohort of patients was highly challenging (92% grade III/IV and only 8% grade II GvHD), after treatment with MEPs the overall survival rate at 2 years follow-up was 71±11% for the entire patient cohort, compared to 51.4±9.0% in GvHD clinical studies, in which MSCs were derived from single donors.
Conclusion
To our knowledge this is the first serum-free MSC bank generated from pooled BM-MNCs of multiple donors as a source for bulk production of clinical-grade MSCs with a predictable potency. Importantly, clinical data presented in this study demonstrated the in vivo safety and efficacy of MEPs, which may provide a novel therapeutic tool for the effective treatment of severe aGVHD.
Session topic: Gene therapy, cellular immunotherapy and vaccination
Keyword(s): Graft-versus-host disease (GVHD), Mesenchymal stem cell
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:45 - 17:00
Location: Hall C14
Background
Mesenchymal stromal cells (MSCs) represent a heterogeneous cell population concerning their proliferative, differentiation and allosuppressive potential.
Aims
To circumvent donor-to-donor heterogeneity which may lead to inconsistent results after treatment of acute graft-versus-host disease (aGvHD) with mesenchymal stromal cells (MSCs) generated from a single or several individual donors, we developed a novel approach by generating MSCs from pooled bone marrow mononuclear cells (BM-MNCs) of eight healthy “third-party” donors.
Methods
BM-MNCs of 8 healthy “third-party” donors were isolated and frozen separately in cryobags. After thawing, BM-MNCs of 8 donors were pooled and cultured for 14 days in DMEM+5% platelet lysate (PL). Generated MSCs were frozen in 209 cryovials ȧ 1.5x10e6 MSCs, representing the MSC-bank. Several vials were expanded for 12-14 days in DMEM+10% PL till the end of passage 2 and frozen as MSC-end products (MEPs). These products were evaluated for proliferative, differentiation and allosuppressive potential as well as genomic stability. Eighty-one MEPs generated from aliquots of the MSC bank were administered on a compassionate-use basis after approval of the regulatory authorities to 26 patients with steroid-refractory acute GvHD at a target dose of 1–2×106 MSCs/kg BW in 7 transplantation centers.
Results
MEPs exhibited typical MSC phenotype, trilineage differentiation potential and replicative senescence after 13 passages, as determined by increased expression of senescence-related markers such as p21 (CDKN1A) and p16 (CDKN2A). Chromosomal analysis of MEPs demonstrated a normal karyotype, whereas FISH analysis demonstrated a normal diploid pattern, indicating their genomic stability. Importantly, MEPs exerted a significantly higher allosuppressive potential than the mean allosuppressive potential of MSCs generated from the same donors individually. Administration of 81 MEPs to 26 patients with severe steroid-resistant aGvHD in 7 stem cell transplant centers who were refractory to many lines of treatment, induced a 77% overall response at the primary endpoint (day 28). Remarkably, although the cohort of patients was highly challenging (92% grade III/IV and only 8% grade II GvHD), after treatment with MEPs the overall survival rate at 2 years follow-up was 71±11% for the entire patient cohort, compared to 51.4±9.0% in GvHD clinical studies, in which MSCs were derived from single donors.
Conclusion
To our knowledge this is the first serum-free MSC bank generated from pooled BM-MNCs of multiple donors as a source for bulk production of clinical-grade MSCs with a predictable potency. Importantly, clinical data presented in this study demonstrated the in vivo safety and efficacy of MEPs, which may provide a novel therapeutic tool for the effective treatment of severe aGVHD.
Session topic: Gene therapy, cellular immunotherapy and vaccination
Keyword(s): Graft-versus-host disease (GVHD), Mesenchymal stem cell
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