CD40-ligand (CD154) gene therapy for chronic lymphocytic leukemia

Blood ◽  
2000 ◽  
Vol 96 (9) ◽  
pp. 2917-2924 ◽  
Author(s):  
William G. Wierda ◽  
Mark J. Cantwell ◽  
Sandra J. Woods ◽  
Laura Z. Rassenti ◽  
Charles E. Prussak ◽  
...  
Keyword(s):  
Gene Therapy ◽  
T Cell ◽  
Chronic Lymphocytic Leukemia ◽  
De Novo ◽  
Leukemia Cell ◽  
Cd40 Ligand ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells ◽  
Interleukin 12 ◽  
Cell Counts

Abstract Chronic lymphocytic leukemia (CLL) cells can be made to express recombinant CD40-ligand (CD154) by transduction with a replication-defective adenovirus vector (Ad-CD154). Ad-CD154–transduced and bystander leukemia cells become highly effective antigen-presenting cells that can induce CLL-specific autologous cytotoxic T lymphocytes in vitro. This study investigated the immunologic and clinical responses to infusion of autologous Ad-CD154-CLL cells in patients with CLL. After a one-time bolus infusion of autologous Ad-CD154–transduced leukemia cells, there was increased or de novo expression of immune accessory molecules on bystander, noninfected CLL cells in vivo. Treated patients also developed high plasma levels of interleukin-12 and interferon-γ, the magnitudes of which corresponded to absolute blood CD4+T-cell counts before therapy. On average, patients experienced a greater than 240% increase in absolute blood T-cell counts within 1 to 4 weeks of treatment. Moreover, treatment increased the numbers of leukemia-specific T cells, demonstrated by autologous ELISPOT assay and mixed lymphocyte reactions. These biologic effects were associated with reductions in leukemia cell counts and lymph node size. Treatment did not induce autoimmune thrombocytopenia or hemolytic anemia and no dose-limiting toxicity was observed. This approach may provide a novel and effective form of gene therapy for patients with this disease.

CD40-ligand (CD154) gene therapy for chronic lymphocytic leukemia

Blood ◽  
2000 ◽  
Vol 96 (9) ◽  
pp. 2917-2924 ◽  
Author(s):  
William G. Wierda ◽  
Mark J. Cantwell ◽  
Sandra J. Woods ◽  
Laura Z. Rassenti ◽  
Charles E. Prussak ◽  
...  
Keyword(s):  
Gene Therapy ◽  
T Cell ◽  
Chronic Lymphocytic Leukemia ◽  
De Novo ◽  
Leukemia Cell ◽  
Cd40 Ligand ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells ◽  
Interleukin 12 ◽  
Cell Counts

Chronic lymphocytic leukemia (CLL) cells can be made to express recombinant CD40-ligand (CD154) by transduction with a replication-defective adenovirus vector (Ad-CD154). Ad-CD154–transduced and bystander leukemia cells become highly effective antigen-presenting cells that can induce CLL-specific autologous cytotoxic T lymphocytes in vitro. This study investigated the immunologic and clinical responses to infusion of autologous Ad-CD154-CLL cells in patients with CLL. After a one-time bolus infusion of autologous Ad-CD154–transduced leukemia cells, there was increased or de novo expression of immune accessory molecules on bystander, noninfected CLL cells in vivo. Treated patients also developed high plasma levels of interleukin-12 and interferon-γ, the magnitudes of which corresponded to absolute blood CD4+T-cell counts before therapy. On average, patients experienced a greater than 240% increase in absolute blood T-cell counts within 1 to 4 weeks of treatment. Moreover, treatment increased the numbers of leukemia-specific T cells, demonstrated by autologous ELISPOT assay and mixed lymphocyte reactions. These biologic effects were associated with reductions in leukemia cell counts and lymph node size. Treatment did not induce autoimmune thrombocytopenia or hemolytic anemia and no dose-limiting toxicity was observed. This approach may provide a novel and effective form of gene therapy for patients with this disease.

scholarly journals CD154 induces p73 to overcome the resistance to apoptosis of chronic lymphocytic leukemia cells lacking functional p53

Blood ◽  
2006 ◽  
Vol 108 (10) ◽  
pp. 3450-3457 ◽  
Author(s):  
Frank Dicker ◽  
Arnon P. Kater ◽  
Carlos E. Prada ◽  
Tetsuya Fukuda ◽  
Januario E. Castro ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Leukemia Cell ◽  
Death Receptor ◽  
Cd40 Ligand ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells ◽  
Cd40 Ligation ◽  
Induced Expression ◽  
Cell Counts ◽  
Induced Apoptosis

Abstract Intravenous infusion of autologous chronic lymphocytic leukemia (CLL) cells transduced with an adenovirus encoding CD40-ligand (CD154) caused rapid reductions in leukemia-cell counts and lymphnode size. We hypothesized that CD40-ligation via CD154 sensitized CLL cells to death-receptor-mediated apoptosis. We found that CD154-expressing cells induced expression of CD95 and the BH3-interacting-domain death agonist (Bid) in CLL, regardless of whether the leukemia cells had functional p53. Such treatment also induced p73, a p53-related transcription factor regulated by c-Abl kinase, and enhanced the sensitivity to fludarabine (F-ara-A) of CLL cells lacking functional p53. Transduction of CLL cells with an adenovirus encoding p73 also induced Bid and CD95 and enhanced the sensitivity to F-ara-A of p53-deficient CLL cells. However, inhibition of c-Abl with imatinib suppressed CD154-induced expression of p73, p73-induced expression of Bid and CD95, and blocked the sensitization of p53-deficient CLL cells to CD95-mediated or F-ara-A-induced apoptosis. Conversely, CLL cells transduced with an imatinib-resistant c-Abl mutant could be induced by CD154 to express p73 and Bid even when treated with imatinib. These results indicate that CD154 can sensitize leukemia cells to apoptosis via the c-Abl-dependent activation of p73 and mitigate the resistance of p53-deficient CLL cells to anticancer drug therapy.

A Phase II, Open Label Study of AT-101 in Combination with Rituximab in Patients with Relapsed or Refractory Chronic Lymphocytic Leukemia.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2838-2838 ◽  
Author(s):  
Januario E. Castro ◽  
Loria J. Olivier ◽  
Aguillon A. Robier ◽  
James Danelle ◽  
Suarez J. Carlos ◽  
...  
Keyword(s):  
High Risk ◽  
Chronic Lymphocytic Leukemia ◽  
Adverse Events ◽  
Plasma Concentrations ◽  
Leukemia Cell ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells ◽  
Pharmacokinetic Studies ◽  
Cell Counts ◽  
Grade 3

Abstract Chronic lymphocytic leukemia (CLL) cells express high-levels of Bcl-2 and related anti-apoptotic proteins that collectively can enhance leukemia-cell survival and drug-resistance. AT-101 is an orally active BH3-mimetic that can inhibit the anti-apoptotic activity of Bcl-2-family-member proteins (e.g. Bcl-2, Bcl-XL, Mcl-1) and induce CLL cells to undergo apoptosis. Furthermore, we found that AT-101 also can enhance the cytotoxicity of rituximab for CLL cells in vitro. As such we conducted a phase 2 trial to evaluate the safety and activity of AT-101 when used together with rituximab to treat 12 patients who had relapsed/refractory CLL. Patients received AT-101, at 30 mg/d, for 21 or 28 days during each of three 28-day cycles. Rituximab was administered at 375 mg/m2 for 12 doses (total dose = 4,500 mg/m2) on days 1, 3, 5, 8, 15, 22, 29, 31, 33, 40, 57, 59, 61. The first dose of rituximab was given over two days to minimize infusion-related adverse events. The patients’ median age was 61.5 years (range 43–81). Nine patients were high risk and 3 were intermediate risk based on the modified Rai classification and had received a median of 2 prior regimens (range 1–8). Six patients had leukemia cells that expressed ZAP-70 and/or unmutated immunoglobulin variable region genes and 4 patients had either 11q deletions or complex cytogenetics. Six patients interrupted treatment due to adverse events, most of which were transient and without residual complications. Grade 1–2 gastrointestinal effects (e.g., nausea, vomiting) occurred in 11 patients, 2 of whom had grade 3/4 ileus. Six patients experienced treatment-associated fatigue (grade 1–2 in 5 and grade 3 in one). Other than ileus and fatigue the only grade 3/4 event noted was neutropenia. One patient without neutropenia died while undergoing treatment from community-acquired bacterial pneumonia[j1]. Pharmacokinetic studies demonstrated that the average Cmax of AT-101 was 565 ng/ml (280 – 805 ng/ml) at a Tmax of 3.1 hours (1.7 – 5.6 hrs.). Correlative science studies performed on leukemia cells isolated at various times after treatment demonstrated leukemia-cell apoptosis in vivo, with maximum levels seen at times when we observed peak drug levels of AT-101. Eight patients had completed the study and had full response evaluation at the time of this abstract’s submission. The overall response rate was 38% [CRu (2); PR (1); SD (3); PD (2)]. Four of eight patients (50%) had significant reductions in leukemia cell counts and splenomegaly and 5 of 8 (63%) had reductions in lymphadenopathy. AT-101 in combination with Rituximab has apparent activity in patients with relapsed-refractory high-risk CLL. Additional enrollment is planned using alternate AT-101 schedules in an attempt to increase peak plasma concentrations (and potentially activity) and reduce GI toxicity.

scholarly journals Chronic lymphocytic leukemia cells induce defective LFA-1–directed T-cell motility by altering Rho GTPase signaling that is reversible with lenalidomide

Blood ◽  
2013 ◽  
Vol 121 (14) ◽  
pp. 2704-2714 ◽  
Author(s):  
Alan G. Ramsay ◽  
Rachel Evans ◽  
Shahryar Kiaii ◽  
Lena Svensson ◽  
Nancy Hogg ◽  
...  
Keyword(s):  
T Cell ◽  
Chronic Lymphocytic Leukemia ◽  
Cell Motility ◽  
Rho Gtpase ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells ◽  
Key Points ◽  
Gtpase Activation ◽  
Activation Signaling

Key Points CLL cells induce defects in T-cell LFA-1–mediated migration by altering Rho GTPase activation signaling, downregulating RhoA and Rac1, and upregulating Cdc42. Lenalidomide repairs these T-cell defects by restoring normal Rho GTPase activation signaling.

scholarly journals A case of chronic lymphocytic leukemia with properties characteristic of natural killer cells

Blood ◽  
1983 ◽  
Vol 61 (5) ◽  
pp. 940-948 ◽  
Author(s):  
K Itoh ◽  
K Tsuchikawa ◽  
T Awataguchi ◽  
K Shiiba ◽  
K Kumagai
Keyword(s):  
T Cell ◽  
Chronic Lymphocytic Leukemia ◽  
Nk Cells ◽  
Natural Killer ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
Leukemia Cells ◽  
Surface Markers ◽  
T Antigens ◽  
Homogeneous Population

Abstract A case of chronic lymphocytic leukemia that consisted of a homogeneous population of cells that had properties similar to those described for natural killer (NK) cells is presented. These leukemic cells had a morphology of large granular lymphocytes (LGL) and receptors for sheep erythrocytes (ER) and for the Fc portion of IgG (Fc gamma-R). They expressed pan-T antigens OKT3 and Leu-4, but neither helper/inducer T- cell differentiation antigens OKT4 and Leu-3a nor cytotoxic/suppressor T-antigens OKT8 and Leu-2a. HNK 1 antigen, which can be expressed on human NK cells, could be detected on almost all leukemic cells (LGL), whereas a myeloid differentiation antigen, OKM1, which can be expressed on macrophages, granulocytes, and NK cells, was not detected. Thus, it was concluded that the leukemia cells had a characteristic profile of the surface markers: ER+, Fc gamma-R+, HNK-1+, OKT3+, Leu-4+, OKT4-, OKT8-, Leu-3a, Leu-2a, and OKM1-. Although freshly isolated leukemic cells showed no cytotoxicity on NK targets, after incubation at 37 degrees C, the cells did show a potent cytotoxicity on targets of erythroleukemic cell, T cell, and monocyte (but not B cell) origins. When the cells were incubated at 37 degrees C, interferon (IFN gamma) was spontaneously produced in the culture fluids. Treatment with anti- HNK-1 and complement completely abrogated expression of NK activity and interferon production of the patient's lymphocytes in culture. These characteristic features of surface markers and functions strongly suggest the possibility that the leukemia cells of this case are of NK cell origin. The relationship between this case and chronic lymphocytic leukemia of T-cell origin is discussed.

Chronic lymphocytic leukemia of Eμ-TCL1 transgenic mice undergoes rapid cell turnover that can be offset by extrinsic CD257 to accelerate disease progression

Blood ◽  
2009 ◽  
Vol 114 (20) ◽  
pp. 4469-4476 ◽  
Author(s):  
Thomas Enzler ◽  
Arnon P. Kater ◽  
Weizhou Zhang ◽  
George F. Widhopf ◽  
Han-Yu Chuang ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Disease Progression ◽  
Leukemia Cell ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells ◽  
Cell Turnover ◽  
Proliferating Cells ◽  
Rapid Reduction ◽  
Disease Evolution ◽  
Dying Cells

AbstractResults of heavy-water labeling studies have challenged the notion that chronic lymphocytic leukemia (CLL) represents an accumulation of noncycling B cells. We examined leukemia cell turnover in Eμ-TCL1 transgenic (TCL1-Tg) mice, which develop a CLL-like disease at 8 to 12 months of age. We found that leukemia cells in these mice not only had higher proportions of proliferating cells but also apoptotic cells than did nonleukemic lymphocytes. We crossed TCL1-Tg with BAFF-Tg mice, which express high levels of CD257. TCL1×BAFF-Tg mice developed CLL-like disease at a significantly younger age and had more rapid disease progression and shorter survival than TCL1-Tg mice. Leukemia cells of TCL1×BAFF-Tg mice had similar proportions of proliferating cells, but fewer proportions of dying cells, than did the CLL cells of TCL1-Tg mice. Moreover, leukemia cells from either TCL1×BAFF-Tg or TCL1-Tg mice produced more aggressive disease when transferred into BAFF-Tg mice than into wild-type (WT) mice. Neutralization of CD257 resulted in rapid reduction in circulating leukemia cells. These results indicate that the leukemia cells of TCL1-Tg mice undergo high levels of spontaneous apoptosis that is offset by relatively high rates of leukemia cell proliferation, which might allow for acquisition of mutations that contribute to disease evolution.

Bystander Transfer of Functional Human CD40 Ligand from Gene-Modified Fibroblasts to B-Chronic Lymphocytic Leukemia Cells

2003 ◽  
Vol 14 (6) ◽  
pp. 545-559 ◽  
Author(s):  
Ettore Biagi ◽  
Eric Yvon ◽  
Gianpietro Dotti ◽  
Persis J. Amrolia ◽  
Satoshi Takahashi ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Cd40 Ligand ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells

CD40-Activation of Chronic Lymphocytic Leukemia Cells Induces Latent Sensitivity to Fas/TRAIL-Mediated Apoptosis Via a P53-Independent, C-Abl-Dependent Pathway.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 342-342 ◽  
Author(s):  
Arnon P. Kater ◽  
Frank Dicker ◽  
Tetsuya Fukuda ◽  
Thomas J. Kipps
Keyword(s):  
Gene Therapy ◽  
Imatinib Mesylate ◽  
Leukemia Cell ◽  
Lymphocytic Leukemia ◽  
Leukemia Cells ◽  
Cd40 Ligation ◽  
Γ Radiation ◽  
Abl Kinase ◽  
Cd40 Activation ◽  
Inactivating Mutations

Abstract Patients with chronic lymphocytic leukemia (CLL) experienced rapid reductions in leukemia cell count and lymph node size following an intravenous infusion of autologous CLL cells that were transduced to express a recombinant ligand for CD40 (CD154), including patients with leukemia cells that had inactivating mutations in P53 . Subsequent studies demonstrated that ligation of CD40 on CLL cells induced latent sensitivity to Fas (CD95)-dependent apoptosis and that this might account, in part, for the early reductions in leukemia cell counts observed in patients treated with CD154 gene therapy (Proc Natl Acad Sci U S A 100:3854–9, 2002). We found that ligation of CD40 induces CLL cells to express P53, as well as P53-dependent genes encoding CD95, DR5, and p21. Similar effects were observed following treatment of CLL cells with γ-radiation. In contrast to the effects of γ-radiation, however, CD40-ligation also induced CLL cells to express high-levels of Bid, a pro-apoptotic protein that mediates cross talk between extrinsic death receptors and mitochondrial-dependent apoptotic pathways. CLL cells that had inactivating mutations in P53 also were induced to express CD95, DR5, p21, and Bid following CD40-ligation, but not following γ-radiation, demonstrating that the gene expression response to CD40-ligation is dependent on a factor(s) other than p53. A good candidate for this is P73, a member of the P53 gene family that has been implicated in lymphocyte activation-induced cell death. Consistent with this notion, we found that ligation of CD40 on CLL cells induced expression of the α isoform of P73. The induction of P73 and latent sensitivity to CD95 (and TRAIL)-mediated apoptosis could be inhibited with the c-abl kinase inhibitor, imatinib mesylate, when added at the time of CD40-ligation. However, transduction of leukemia cells with a mutant gene encoding a c-abl kinase resistant to imatinib mesylate resulted in CLL cells that could be induced by CD40-ligation to express P73 and Bid even in the presence of imatinib mesylate. We conclude that sensitization to CD95-mediated apoptosis following CD40-activation is via a p53-independent, c-abl-kinase dependent process that is associated with activation of p73. Unlike p53, p73 mutations are extremely rare in human cancer. As such, CD154-based immune gene therapy may have activity even in patients who have CLL cells with dysfunctional P53 that are resistant to many of the anti-leukemia drugs currently used in the treatment of this disease.

Lenalidomide Abrogates the Protective Influence of Nurse-Like Cells on Primary Chronic Lymphocytic Leukemia Cells In Vitro.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3116-3116 ◽  
Author(s):  
Danelle F. James ◽  
Maryann R. Betty ◽  
Ruzbeh Mosadeghi ◽  
Thomas J. Kipps
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Cell Viability ◽  
Cell Survival ◽  
Leukemia Cell ◽  
Lymphocytic Leukemia ◽  
Response To Treatment ◽  
Leukemia Cells ◽  
High Dependency ◽  
Cells Cultured

Abstract Lenalidomide (3-(4-amino-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione)) is an agent approved for treatment of patients with del 5q myelodysplastic syndromes and previously treated multiple myeloma. Lenalidomide has been found in early clinical trials to have potential therapeutic activity in patients with relapsed chronic lymphocytic leukemia (CLL). The mechanism(s) whereby this drug is active in CLL is unknown. In particular, studies to date have not found lenalidomide to have any direct cytotoxic activity on CLL cells in vitro. This has stimulated speculation that this agent might adversely affect the positive influence of the microenvironment on leukemia-cell survival. We and others have observed that cells found in the leukemia microenvironment can support CLL-cell survival in vitro. One such type of cells are nurse-like cells (NLC), which can differentiate from the CD14-positive blood mononuclear cells of CLL patients into large, round adherent cells that can attract and support CLL cell survival in vitro for weeks, if not longer. We evaluated the effects of lenalidomide on primary leukemia-cell survival in vitro when the CLL cells from different patients (N=21) were cultured alone or together with NLC generated as previously described [Tsukada Blood 2002]. We assessed the in-vitro activity of lenalidomide on primary CLL cells from 21 patients, in duplicate in a series of 6 experiments. Lenalidomide at concentrations of 0.1μM-200μM did not significantly impact the survival of CLL cells that were cultured alone for up to 12 days. Analysis of cell surface markers revealed increased expression of CD38 at 36 hours in 5/5 lenalidomide treated CLL samples compared with untreated cells (MFIR 5.7 +/− .86 vs. 3.4 +/− .83 p=.003). We observed sustained upregualtion of CD40 and regulation of CXCR4 in the majority of cells treated with lenalidomide. When cultured with NLC, the survival of CLL cells was comparable to or significantly higher than that of CLL cells cultured alone 62.4% vs. 51% (+/−3% SEM n=21 p [<] 0.0005). The addition of lenalidomide at concentrations of 0.1μM and greater to co-cultures of NLC and CLL cells caused specific reductions in CLL cell survival to levels similar to or lower than that of CLL cells cultured without NLC. In the presence of NLC, lenalidomide at 1μM reduced CLL cell viability compared to control (41.5% vs. 56% +/−4% p [<] 0.0005 paired student t test n=13). For most patients the levels of CLL cell viability on days 4 through 8 in the co-cultures with lenalidomide was significantly lower than those of CLL cells co-cultured with NLC in the absence of lenalidomide. As such, this study reveals that physiologic concentrations of lenalidomide might abrogate the protective influence of NLC on CLL cell survival in vitro and potentially in vivo. Conceivably, those patients who have leukemia cells displaying a high dependency on NLC for survival in vitro also might be most likely to experience a favorable clinical response to treatment with lenalidomide. This hypothesis will be tested in a prospective manner with a planned clinical trial evaluating lenalidomide for treatment of CLL through the CLL Research Consortium.

Sign in / Sign up

Export Citation Format

Share Document