DONOR-DERIVED, CD19-DIRECTED, CAR-MODIFIED T CELLS INFUSED AFTER ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION AS PRE-EMPTIVE DONOR LYMPHOCYTE INFUSION IN PATIENTS WITH CD19+ MALIGNANCIES
(Abstract release date: 05/21/15)
EHA Library. Kebriaei P. 06/14/15; 103162; S802
Disclosure(s): MD Anderson Cancer Center
Dr. Partow Kebriaei
Contributions
Contributions
Abstract
Abstract: S802
Type: Oral Presentation
Presentation during EHA20: From 14.06.2015 08:30 to 14.06.2015 08:45
Location: Room Lehar 1 + 2
Background
Allogeneic hematopoietic stem cell transplantation (HSCT) can be curative in a subset of patients with advanced B-lineage acute lymphoblastic leukemia (ALL), but relapse remains the main reason for treatment failure. Donor-derived, non-specific lymphocyte infusions (DLI) have been ineffectively infused and are associated with significant graft-versus-host-disease (GVHD). Chimeric antigen receptor (CAR)-modified T cells directed toward CD19 have demonstrated dramatic efficacy in patients with refractory ALL. However, responses are often associated with life-threatening cytokine release.
Aims
We hypothesized that infusing CAR-modified, CD19-specific T cells after HSCT as a directed DLI would be associated with less GVHD while providing cellular-based disease control, and may be associated with less cytokine released when administered in a minimal disease state.
Methods
We employed a non-viral gene transfer using the Sleeping Beauty (SB) transposon/transposase system to stably express a 2nd generation CD19-specific CAR (designated CD19RCD28 that activates via CD3z/CD28) in donor-derived T cells for patients with advanced CD19+ lymphoid malignancies. T cells were electroporated using a Nucleofector device to synchronously introduce two DNA plasmids coding for SB transposon (CD19RCD28) and hyperactive SB transposase (SB11). T cells stably expressing the CAR were retrieved over 28 days of co-culture by recursive additions of g-irradiated activating and propagating cells (AaPC) in presence of soluble recombinant interleukin (IL)-2 and IL-21. The AaPC were derived from K562 cells and genetically modified to co-express CD19 as well as the co-stimulatory molecules CD86, CD137L, and a membrane-bound version of IL-15.
Results
To date, we have successfully treated 16 patients with advanced CD19+ ALL (n=13) or NHL (n=3); 7 patients had active disease at time of HSCT. Donor-derived CAR+ T cells (HLA-matched sibling n=9; haplo-family n=5; double cord blood n=2) were infused at a median 64 days (range 42-91 days) following HSCT to prevent disease progression. Transplant preparative regimens were myeloablative, busulfan-based (n=8) or reduced intensity, fludarabine-based (n=8). All patients were maintained on GVHD prophylaxis at time of CAR infusion with tacrolimus, plus mycophenolate mofeteil for cord, plus post-HSCT cyclophosphamide for haplo donors. The starting CAR+ T-cell dose was 106 (n=7), escalated to 107 (n=6), and currently at 5x107 (n=3) modified T cells/m2 (based on recipient body surface area). Patients have not demonstrated any acute or late toxicity to CAR+ T cell infusions. Three patients developed acute grades 2-4 GVHD (liver n=1, upper GI n=1, skin=1) which was within the expected range after allogeneic HSCT. Eight patients have relapsed at a median of 90 days following HCT (range 68-185 days). Fifty percent of patients (n=8) remain alive and in complete remission (CR) at median 7.2 months (range 2.1-21.3 months) following HSCT.
Summary
Keyword(s): Acute lymphoblastic leukemia, Adoptive immunotherapy, Allogeneic hematopoietic stem cell transplant
Session topic: Stem cell transplantation: Clinical 3
Type: Oral Presentation
Presentation during EHA20: From 14.06.2015 08:30 to 14.06.2015 08:45
Location: Room Lehar 1 + 2
Background
Allogeneic hematopoietic stem cell transplantation (HSCT) can be curative in a subset of patients with advanced B-lineage acute lymphoblastic leukemia (ALL), but relapse remains the main reason for treatment failure. Donor-derived, non-specific lymphocyte infusions (DLI) have been ineffectively infused and are associated with significant graft-versus-host-disease (GVHD). Chimeric antigen receptor (CAR)-modified T cells directed toward CD19 have demonstrated dramatic efficacy in patients with refractory ALL. However, responses are often associated with life-threatening cytokine release.
Aims
We hypothesized that infusing CAR-modified, CD19-specific T cells after HSCT as a directed DLI would be associated with less GVHD while providing cellular-based disease control, and may be associated with less cytokine released when administered in a minimal disease state.
Methods
We employed a non-viral gene transfer using the Sleeping Beauty (SB) transposon/transposase system to stably express a 2nd generation CD19-specific CAR (designated CD19RCD28 that activates via CD3z/CD28) in donor-derived T cells for patients with advanced CD19+ lymphoid malignancies. T cells were electroporated using a Nucleofector device to synchronously introduce two DNA plasmids coding for SB transposon (CD19RCD28) and hyperactive SB transposase (SB11). T cells stably expressing the CAR were retrieved over 28 days of co-culture by recursive additions of g-irradiated activating and propagating cells (AaPC) in presence of soluble recombinant interleukin (IL)-2 and IL-21. The AaPC were derived from K562 cells and genetically modified to co-express CD19 as well as the co-stimulatory molecules CD86, CD137L, and a membrane-bound version of IL-15.
Results
To date, we have successfully treated 16 patients with advanced CD19+ ALL (n=13) or NHL (n=3); 7 patients had active disease at time of HSCT. Donor-derived CAR+ T cells (HLA-matched sibling n=9; haplo-family n=5; double cord blood n=2) were infused at a median 64 days (range 42-91 days) following HSCT to prevent disease progression. Transplant preparative regimens were myeloablative, busulfan-based (n=8) or reduced intensity, fludarabine-based (n=8). All patients were maintained on GVHD prophylaxis at time of CAR infusion with tacrolimus, plus mycophenolate mofeteil for cord, plus post-HSCT cyclophosphamide for haplo donors. The starting CAR+ T-cell dose was 106 (n=7), escalated to 107 (n=6), and currently at 5x107 (n=3) modified T cells/m2 (based on recipient body surface area). Patients have not demonstrated any acute or late toxicity to CAR+ T cell infusions. Three patients developed acute grades 2-4 GVHD (liver n=1, upper GI n=1, skin=1) which was within the expected range after allogeneic HSCT. Eight patients have relapsed at a median of 90 days following HCT (range 68-185 days). Fifty percent of patients (n=8) remain alive and in complete remission (CR) at median 7.2 months (range 2.1-21.3 months) following HSCT.
Summary
We report the first human application of the SB and AaPC platforms to genetically modify clinical-grade cells. We demonstrate that infusing donor-derived CD19-specific CAR+ T cells in the adjuvant HSCT setting as pre-emptive DLI may provide an effective and safe approach for maintaining remission in patients at high risk for relapse. Modification of the CAR construct is underway in efforts to improve CAR T-cell in vivo proliferation and persistence.
Keyword(s): Acute lymphoblastic leukemia, Adoptive immunotherapy, Allogeneic hematopoietic stem cell transplant
Session topic: Stem cell transplantation: Clinical 3
Abstract: S802
Type: Oral Presentation
Presentation during EHA20: From 14.06.2015 08:30 to 14.06.2015 08:45
Location: Room Lehar 1 + 2
Background
Allogeneic hematopoietic stem cell transplantation (HSCT) can be curative in a subset of patients with advanced B-lineage acute lymphoblastic leukemia (ALL), but relapse remains the main reason for treatment failure. Donor-derived, non-specific lymphocyte infusions (DLI) have been ineffectively infused and are associated with significant graft-versus-host-disease (GVHD). Chimeric antigen receptor (CAR)-modified T cells directed toward CD19 have demonstrated dramatic efficacy in patients with refractory ALL. However, responses are often associated with life-threatening cytokine release.
Aims
We hypothesized that infusing CAR-modified, CD19-specific T cells after HSCT as a directed DLI would be associated with less GVHD while providing cellular-based disease control, and may be associated with less cytokine released when administered in a minimal disease state.
Methods
We employed a non-viral gene transfer using the Sleeping Beauty (SB) transposon/transposase system to stably express a 2nd generation CD19-specific CAR (designated CD19RCD28 that activates via CD3z/CD28) in donor-derived T cells for patients with advanced CD19+ lymphoid malignancies. T cells were electroporated using a Nucleofector device to synchronously introduce two DNA plasmids coding for SB transposon (CD19RCD28) and hyperactive SB transposase (SB11). T cells stably expressing the CAR were retrieved over 28 days of co-culture by recursive additions of g-irradiated activating and propagating cells (AaPC) in presence of soluble recombinant interleukin (IL)-2 and IL-21. The AaPC were derived from K562 cells and genetically modified to co-express CD19 as well as the co-stimulatory molecules CD86, CD137L, and a membrane-bound version of IL-15.
Results
To date, we have successfully treated 16 patients with advanced CD19+ ALL (n=13) or NHL (n=3); 7 patients had active disease at time of HSCT. Donor-derived CAR+ T cells (HLA-matched sibling n=9; haplo-family n=5; double cord blood n=2) were infused at a median 64 days (range 42-91 days) following HSCT to prevent disease progression. Transplant preparative regimens were myeloablative, busulfan-based (n=8) or reduced intensity, fludarabine-based (n=8). All patients were maintained on GVHD prophylaxis at time of CAR infusion with tacrolimus, plus mycophenolate mofeteil for cord, plus post-HSCT cyclophosphamide for haplo donors. The starting CAR+ T-cell dose was 106 (n=7), escalated to 107 (n=6), and currently at 5x107 (n=3) modified T cells/m2 (based on recipient body surface area). Patients have not demonstrated any acute or late toxicity to CAR+ T cell infusions. Three patients developed acute grades 2-4 GVHD (liver n=1, upper GI n=1, skin=1) which was within the expected range after allogeneic HSCT. Eight patients have relapsed at a median of 90 days following HCT (range 68-185 days). Fifty percent of patients (n=8) remain alive and in complete remission (CR) at median 7.2 months (range 2.1-21.3 months) following HSCT.
Summary
Keyword(s): Acute lymphoblastic leukemia, Adoptive immunotherapy, Allogeneic hematopoietic stem cell transplant
Session topic: Stem cell transplantation: Clinical 3
Type: Oral Presentation
Presentation during EHA20: From 14.06.2015 08:30 to 14.06.2015 08:45
Location: Room Lehar 1 + 2
Background
Allogeneic hematopoietic stem cell transplantation (HSCT) can be curative in a subset of patients with advanced B-lineage acute lymphoblastic leukemia (ALL), but relapse remains the main reason for treatment failure. Donor-derived, non-specific lymphocyte infusions (DLI) have been ineffectively infused and are associated with significant graft-versus-host-disease (GVHD). Chimeric antigen receptor (CAR)-modified T cells directed toward CD19 have demonstrated dramatic efficacy in patients with refractory ALL. However, responses are often associated with life-threatening cytokine release.
Aims
We hypothesized that infusing CAR-modified, CD19-specific T cells after HSCT as a directed DLI would be associated with less GVHD while providing cellular-based disease control, and may be associated with less cytokine released when administered in a minimal disease state.
Methods
We employed a non-viral gene transfer using the Sleeping Beauty (SB) transposon/transposase system to stably express a 2nd generation CD19-specific CAR (designated CD19RCD28 that activates via CD3z/CD28) in donor-derived T cells for patients with advanced CD19+ lymphoid malignancies. T cells were electroporated using a Nucleofector device to synchronously introduce two DNA plasmids coding for SB transposon (CD19RCD28) and hyperactive SB transposase (SB11). T cells stably expressing the CAR were retrieved over 28 days of co-culture by recursive additions of g-irradiated activating and propagating cells (AaPC) in presence of soluble recombinant interleukin (IL)-2 and IL-21. The AaPC were derived from K562 cells and genetically modified to co-express CD19 as well as the co-stimulatory molecules CD86, CD137L, and a membrane-bound version of IL-15.
Results
To date, we have successfully treated 16 patients with advanced CD19+ ALL (n=13) or NHL (n=3); 7 patients had active disease at time of HSCT. Donor-derived CAR+ T cells (HLA-matched sibling n=9; haplo-family n=5; double cord blood n=2) were infused at a median 64 days (range 42-91 days) following HSCT to prevent disease progression. Transplant preparative regimens were myeloablative, busulfan-based (n=8) or reduced intensity, fludarabine-based (n=8). All patients were maintained on GVHD prophylaxis at time of CAR infusion with tacrolimus, plus mycophenolate mofeteil for cord, plus post-HSCT cyclophosphamide for haplo donors. The starting CAR+ T-cell dose was 106 (n=7), escalated to 107 (n=6), and currently at 5x107 (n=3) modified T cells/m2 (based on recipient body surface area). Patients have not demonstrated any acute or late toxicity to CAR+ T cell infusions. Three patients developed acute grades 2-4 GVHD (liver n=1, upper GI n=1, skin=1) which was within the expected range after allogeneic HSCT. Eight patients have relapsed at a median of 90 days following HCT (range 68-185 days). Fifty percent of patients (n=8) remain alive and in complete remission (CR) at median 7.2 months (range 2.1-21.3 months) following HSCT.
Summary
We report the first human application of the SB and AaPC platforms to genetically modify clinical-grade cells. We demonstrate that infusing donor-derived CD19-specific CAR+ T cells in the adjuvant HSCT setting as pre-emptive DLI may provide an effective and safe approach for maintaining remission in patients at high risk for relapse. Modification of the CAR construct is underway in efforts to improve CAR T-cell in vivo proliferation and persistence.
Keyword(s): Acute lymphoblastic leukemia, Adoptive immunotherapy, Allogeneic hematopoietic stem cell transplant
Session topic: Stem cell transplantation: Clinical 3
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