Alan S Wayne

Children's Hospital Los Angeles, Los Angeles, California, United States

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Publications (78)348.97 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: NK cells can enhance engraftment and mediate GVL following allogeneic HSCT, but the potency of GVL mediated by naturally reconstituting NK cells following HSCT is limited. Preclinical studies demonstrate that activation of NK cells using IL-15 plus 4-1BBL upregulates activating receptor expression and augments killing capacity. In an effort to amplify the beneficial effects of NK cells post-HSCT, we conducted a first-in-human trial of adoptive transfer of donor-derived IL-15/4-1BBL activated NK cells (aNK-DLI) following HLA-matched, T cell depleted (1-2 x 10(4) T cells/kg) non-myeloablative PBSCT in children and young adults with ultra-high risk solid tumors (Clinicaltrials.gov (NCT01287104)). Activated NK-DLI were CD3(+) depleted, CD56(+) selected lymphocytes, cultured for 9-11 days with rhIL-15 plus 4-1BBL(+)IL-15Rα(+)artificial APCs. aNK-DLI demonstrated potent killing capacity and displayed high levels of activating receptor expression. Five of 9 transplant recipients experienced acute GVHD following aNK-DLI, with Grade 4 GVHD observed in 3 subjects. GVHD was more common in matched unrelated donor vs matched sibling donor recipients, and was associated with higher donor CD3 chimerism. Given that the T cell dose was below the threshold required for GVHD in this setting, we conclude that aNK-DLI contributed to the acute GVHD observed, likely by augmenting underlying T cell alloreactivity. Copyright © 2014 American Society of Hematology.
    Blood 12/2014; · 9.78 Impact Factor
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    ABSTRACT: The diagnostic criteria for paediatric mastocytosis are largely based on adult studies and bone marrow findings are not well described in children. We evaluated use of the World Health Organization (WHO) criteria for the diagnosis of systemic disease in paediatric mastocytosis. In addition, we identified unique clinico-histopathological features within the biopsies. One hundred and thirteen children with paediatric mastocytosis were evaluated at the National Institutes of Health between 1986 and 2013. Complete bone marrow evaluations were performed in 50 cases. Seven children had repeat procedures. Bone marrows were analysed by histopathology, flow cytometry and for KIT D816V. Bone marrow biopsies displayed mild atypical haematopoietic maturation, increased haematogones and hypocellularity in a sub-set of patients with urticaria pigmentosa, diffuse cutaneous mastocytosis and indolent systemic mastocytosis. Hypocellularity was most pronounced in those with urticaria pigmentosa. Haematogones were highest, on average, in patients with diffuse cutaneous mastocytosis or mastocytomas. There was no evidence of peripheral blood cytopenias, myelodysplastic syndrome, myeloproliferative neoplasm or leukaemia within this cohort. The WHO criteria are applicable for the diagnosis of systemic mastocytosis in paediatrics. Although unsuspected bone marrow findings typically seen in myeloproliferative disorders are frequent in paediatric mastocytosis, patients within this study remained clinically stable without progression to a more aggressive variant.
    British Journal of Haematology 11/2014; · 4.94 Impact Factor
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    ABSTRACT: With increasing frequency, allogeneic hematopoietic cell transplantation involving children is being performed in the research setting. Allogeneic hematopoietic cell transplantation, however, cannot be performed without a hematopoietic stem cell (HSC) donor. This donor is often a sibling of the recipient and may also be a child. In such circumstances, it is unclear whether or how the federal regulations for pediatric research apply to the minor donors. This introductory paper reviews the issues to be considered while evaluating studies that use HSCs obtained from minor donors and identifies areas where further research is needed. In the era of increasing applicability for donor-derived cellular therapies, we provide a suggested framework for determining when minor donors qualify as human research subjects and when their participation can be approved under the federal regulations.Bone Marrow Transplantation advance online publication, 20 October 2014; doi:10.1038/bmt.2014.224.
    Bone marrow transplantation. 10/2014;
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    ABSTRACT: Chimeric antigen receptor (CAR) modified T cells targeting CD19 have shown activity in case series of patients with acute and chronic lymphocytic leukaemia and B-cell lymphomas, but feasibility, toxicity, and response rates of consecutively enrolled patients treated with a consistent regimen and assessed on an intention-to-treat basis have not been reported. We aimed to define feasibility, toxicity, maximum tolerated dose, response rate, and biological correlates of response in children and young adults with refractory B-cell malignancies treated with CD19-CAR T cells.
    Lancet. 10/2014;
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    ABSTRACT: Although most children with ALL can be cured by chemotherapy approaches, allogeneic hematopoietic cell transplant (HCT) therapy offers a better chance of cure to selected high-risk patients in first remission and most children who relapse. Although transplant-related mortality has decreased significantly in the past decade, relapse remains high after HCT for ALL; developing strategies to decrease relapse and improve survival are vital. Recent studies have shown that relapse risk can be accurately defined using measurements of minimal residual disease (MRD) both pre- and post-HCT and by knowing whether patients get GVHD in the first 2 months after transplant. With these risk definitions in hand, investigators are now applying novel agents and immunotherapeutic methods in attempt to lower MRD before transplant and modulate the GVL effect after transplant. With powerful new immunological approaches coming on line, the transplant process itself will likely expand to include pre and/or post-HCT interventions aimed at reducing relapse.Bone Marrow Transplantation advance online publication, 16 June 2014; doi:10.1038/bmt.2014.114.
    Bone marrow transplantation. 06/2014;
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    ABSTRACT: The presence of minimal residual disease (MRD) pre-transplant is the most important prognostic risk factor predictive of post-transplant relapse in hematologic malignancies. However, MRD alone does not adequately predict relapse in all patients. To improve upon the ability to identify patients likely to relapse, we evaluated risk factors, in addition to MRD, which may be associated with development of post-transplant relapse. In this single institution, retrospective cohort study of children with acute leukemia or myelodysplastic syndrome who had undergone a first allogeneic transplant and had pre-transplant MRD evaluation, 40 of 93 patients (43%) experienced relapse. Univariate analysis demonstrated that African American race, high initial white blood cell count, central nervous system (CNS) disease at diagnosis, short first complete remission, non-myeloablative (NMA) conditioning, lack of remission and MRD pre-transplant were associated with worse relapse-free survival (RFS). In a Cox multivariable analysis, CNS disease (p=0.009), lack of remission (p=0.01) and NMA conditioning (p=0.04) were independently associated with inferior RFS. Amongst those in a morphologic complete remission who underwent a myeloablative transplant, having both CNS disease at diagnosis (specifically in acute lymphoblastic leukemia) and MRD positivity was an independent risk factor predictive of relapse, which has not been previously reported. Results from our study support the existence of risk factors complimentary to pre-transplant MRD. Validation in a larger independent homogenous cohort is needed to develop a prognostic tool for clinical use to predict post-transplant relapse.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 03/2014; · 3.15 Impact Factor
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    ABSTRACT: Outcomes for patients with hematologic malignancies who experience overt relapse after allogeneic hematopoietic stem cell transplantation (HCT) are poor. There are limited data on outcomes of post-transplant minimal residual disease (MRD). In this single institution, retrospective cohort analysis of children with acute leukemia and myelodysplastic syndrome we document the pattern of relapse with a primary focus on outcomes of post-transplant MRD. Forty of 93 (43%) patients who underwent a first allogeneic HCT and who all had systematic pre and post-transplant MRD evaluations at +30, +60, +90, +180 days and at +1 and +2 years post-transplant, experienced relapse. The median time to relapse was 4.8 months post-transplant with a median survival of 4 months post-relapse. Despite frequent, systematic, routine post-HCT disease restaging evaluation, 31 patients (78%) presented with overt disease at the time of relapse. 7 patients with acute leukemia who had post-transplant MRD, presented at a median of 1 month post-transplant. Due to rapid disease progression or treatment-related mortality (TRM), there was no improvement in survival for those patients whose leukemia was detected in a state of MRD post-transplant. Our results suggest that early intervention strategies targeting post-transplant MRD for relapse prevention in acute leukemia may not be feasible.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 03/2014; · 3.15 Impact Factor
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    ABSTRACT: Unconjugated monoclonal antibodies that target hematopoietic differentiation antigens have been developed to treat hematologic malignancies. Although some of these have activity against chronic lymphocytic leukemia and hairy cell leukemia, in general, monoclonal antibodies have limited efficacy as single agents in the treatment of leukemia. To increase their potency, the binding domains of monoclonal antibodies can be attached to protein toxins. Such compounds, termed immunotoxins, are delivered to the interior of leukemia cells based on antibody specificity for cell surface target antigens. Recombinant immunotoxins have been shown to be highly cytotoxic to leukemic blasts in vitro, in xenograft model systems and in early phase clinical trials in humans. These agents will likely play an increasing role in the treatment of leukemia.
    Blood 02/2014; · 9.78 Impact Factor
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    ABSTRACT: In the National Cancer Institute's Second Workshop on the Biology, Prevention, and Treatment of Relapse After Hematopoietic Stem Cell Transplantation, the Scientific/Educational Session on the Biology of Relapse discussed recent advances in understanding some of the host, disease and transplant-related contributions to relapse, emphasizing concepts with potential therapeutic implications. Relapse after hematopoietic stem cell transplantation (HSCT) represents tumor escape - from the cytotoxic effects of the conditioning regimen and from immunologic control mediated by reconstituted lymphocyte populations. Factors influencing the biology of the therapeutic graft-versus-malignancy (GVM) effect - and relapse - include conditioning regimen effects upon lymphocyte populations and homeostasis, immunologic niches, and the tumor microenvironment; reconstitution of lymphocyte populations and establishment of functional immune competence; and genetic heterogeneity within the malignancy defining potential for clonal escape. Recent developments in T- and NK-cell homeostasis and reconstitution are reviewed, with implications for prevention and treatment of relapse, as is the application of modern genome sequencing to defining the biologic basis of GVM, clonal escape and relapse after HSCT.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 09/2013; · 3.15 Impact Factor
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    ABSTRACT: In the National Cancer Institute's Second International Workshop on the Biology, Prevention, and Treatment of Relapse after Hematopoietic Stem Cell Transplantation, the Scientific/Educational Session on Autologous Transplantation addressed the role of novel agents and immunomodulatory strategies in management of relapse after autologous hematopoietic stem cell transplantation (AHSCT). Concepts were illustrated through in-depth discussion of multiple myeloma, with broader discussion of areas relevant for relapse of other malignancies as well as in the setting of allogeneic transplantation. Dr. Hari provided an overview of the epidemiology of relapse after AHSCT in multiple myeloma, addressing clinical patterns, management implications, and treatment options at relapse, highlighting the implications of novel therapeutic agents in initial, maintenance and relapse treatment. Dr. Avigan discussed current concepts in tumor vaccine design, including whole-cell and antigen-specific strategies, use of an AHSCT platform to reverse tumor-associated immunosuppression and tolerance, and combining vaccines with immunomodulatory agents to promote establishment of durable antitumor immunity. Dr. Hsu reviewed the immunogenetics of natural killer (NK) cells and general NK biology, the clinical importance of autologous NK activity (e.g., lymphoma and neuroblastoma), as well as the impact of existing therapies on promotion of NK-cell activity (e.g., immunomodulatory drugs, monoclonal antibodies) and strategies for enhancing autologous and allogeneic NK-cell effects through NK-cell gene profiling.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 09/2013; · 3.15 Impact Factor
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    ABSTRACT: In the 2(nd) NCI Workshop on the Biology, Prevention, and Treatment of Relapse After Hematopoietic Stem Cell Transplantation, the Scientific/Educational Session on the Prevention and Treatment of Relapse after Allogeneic Transplantation highlighted progress in developing new therapeutic approaches since the 1(st) Relapse Workshop. Recent insights that might provide a basis for the development of novel, practical clinical trials were emphasized, including utilization of newer agents, optimization of donor lymphocyte infusion (DLI), and investigation of novel cellular therapies. Dr. de Lima discussed preemptive and maintenance strategies to prevent relapse after transplantation, e.g., recent promising results suggestive of enhanced graft-versus-tumor activity with hypomethylating agents. Dr. Schmid provided an overview of adjunctive strategies to improve cell therapy for relapse, including cytoreduction prior to DLI, combination of targeted agents with DLI, and considerations in use of second transplants. Dr. Porter addressed strategies to enhance T-cell function, including ex-vivo activated T cells and T-cell engineering, and immunomodulatory approaches to enhance T-cell function in vivo, including exogenous cytokines and modulation of costimulatory pathways.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 09/2013; · 3.15 Impact Factor
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    ABSTRACT: Adoptive immunotherapy with the use of chimeric antigen receptor (CAR)-engineered T cells specific for CD19 has shown promising results for the treatment of B-cell lymphomas and leukemia. This therapy involves the transduction of autologous T cells with a viral vector and the subsequent cell expansion. We describe a new, simplified method to produce anti-CD19-CAR T cells. T cells were isolated from peripheral blood mononuclear cell (PBMC) with anti-CD3/anti-CD28 paramagnetic beads. After 2 days, the T cells were added to culture bags pre-treated with RetroNectin and loaded with the retroviral anti-CD19 CAR vector. The cells, beads and vector were incubated for 24 h, and a second transduction was then performed. No spinoculation was used. Cells were then expanded for an additional 9 days. The method was validated through the use of two PBMC products from a patient with B-cell chronic lymphoblastic leukemia and one PBMC product from a healthy subject. The two PBMC products from the patient with B-cell chronic lymphoblastic leukemia contained 11.4% and 12.9% T cells. The manufacturing process led to final products highly enriched in T cells with a mean CD3+ cell content of 98%, a mean expansion of 10.6-fold and a mean transduction efficiency of 68%. Similar results were obtained from the PBMCs of the first four patients with acute lymphoblastic leukemia treated at our institution. We developed a simplified, semi-closed system for the initial selection, activation, transduction and expansion of T cells with the use of anti-CD3/anti-CD28 beads and bags to produce autologous anti-CD19 CAR-transduced T cells to support an ongoing clinical trial.
    Cytotherapy 08/2013; · 3.06 Impact Factor
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    ABSTRACT: Moxetumomab pasudotox (HA22) is an immunotoxin with an anti-CD22 Fv fused to a portion of Pseudomonas exotoxin A that kills CD22 expressing ALL cells. HA22 produced significant responses in some cases of ALL. To understand how to increase response rate, we isolated HA22-resistant KOPN-8 cells and found that HA22 cannot inactivate elongation factor-2 (EF2) due to low levels of DPH1 RNA and protein. Resistance was associated with methylation of the CpG island in the DPH1 promoter. 5-Azacytidine prevented resistance and methylation of the CpG residues and merits evaluation to determine if it can increase the efficacy of HA22 in ALL.
    Leukemia research 08/2013; · 2.36 Impact Factor
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    ABSTRACT: HA22 is a recombinant immunotoxin that kills CD22 expressing cells by ADP-ribosylating and inactivating elongation factor-2 (EF2). HA22 is composed of an Fv that binds to CD22 that is fused to a potion of Pseudomonas exotoxin A. HA22 is very active in drug resistant Hairy Cell Leukemia, but less active in children with acute lymphoblastic leukemia. To understand why some patients don't respond to HA22, we isolated a HA22 resistant lymphoma cell line and showed that resistance is due to the inability of HA22 to ADP-ribosylate and inactivate EF2. We analyzed the diphthamide synthesis genes and found that WDR85 gene was deleted. We showed that WDR85 knockdown confers HA22 resistance to sensitive cells and sensitivity is restored by introduction of a WDR85 cDNA into resistant cells. Analysis of EF2 in the mutant cells revealed a novel form of diphthamide with an additional methyl group that prevents ADP-ribosylation and inactivation of EF2. The abnormal methylation appears to be catalyzed by DPH5. Inactivation of the WDR85 gene could be a mechanism of immunotoxin resistance in patients undergoing immunotoxin therapy.
    Journal of Biological Chemistry 03/2013; · 4.65 Impact Factor
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    ABSTRACT: There is a paucity of pharmacokinetic studies describing weight-based dosing of intravenous voriconazole in obese patients. In this case report, we describe the pharmacokinetics of intravenous voriconazole in an obese CYP2C19 homozygous poor metabolizer and review previously reported data regarding the use of intravenous voriconazole in obese patients. A 17-year-old obese Hispanic male patient (body mass index 35 kg/m ) received intravenous voriconazole for the treatment of suspected aspergillosis. After 2.5 days of voriconazole 4 mg/kg intravenously every 12 hours based on adjusted body weight, the voriconazole area under the serum concentration-time curve over the course of a single (12-hr) dosing interval and trough concentration were 86,100 ng · hr/ml and 6.2 µg/ml, respectively. Six days later, the voriconazole dosage was decreased. A trough concentration measured just before the dosage reduction (after 8.5 days of voriconazole 4 mg/kg intravenously every 12 hours based on adjusted body weight) remained elevated at 5.8 µg/ml. Genotyping revealed a CYP2C19 homozygous poor metabolizer (CYP2C19*2/*2). Voriconazole was subsequently discontinued due to QTc prolongation. These data and those from two recent publications suggest that voriconazole does not distribute extensively into human adipose tissue and that obese patients should be dosed on an adjusted body weight basis. If an obese patient dosed on total body weight is also a CYP2C19 poor metabolizer, serum voriconazole concentrations will be further elevated, potentially leading to drug-induced toxicity.
    Pharmacotherapy 03/2013; 33(3):e19-22. · 2.31 Impact Factor
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    Nirali N Shah, Hema Dave, Alan S Wayne
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    ABSTRACT: Substantial progress has been made in the treatment of leukemia in childhood. Despite this, leukemia remains a leading cause of pediatric cancer-related mortality and the prognosis is guarded for individuals with relapsed or refractory disease. Standard therapies are associated with a wide array of acute and long-term toxicities and further treatment intensification may not be tolerable or beneficial. The curative potential of allogeneic stem cell transplantation is due in part to the graft-versus-leukemia effect, which provides evidence for the therapeutic capacity of immune-based therapies. In recent years there have been significant advances in the development and application of immunotherapy in the treatment of leukemias, including the demonstration of activity in chemotherapy-resistant cases. This review summarizes immunotherapeutic approaches in the treatment of pediatric leukemia including current results and future directions.
    Frontiers in Oncology 01/2013; 3:166.
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    ABSTRACT: Immune targeting of B cell malignancies using chimeric antigen receptors (CARs) is a promising new approach, but critical factors impacting CAR efficacy remain unclear. To test the suitability of targeting CD22 on precursor B cell acute lymphoblastic leukemia (BCP-ALL), lymphoblasts from 111 patients with BCP-ALL were assayed for CD22 expression and all were found to be CD22 positive, with median CD22 expression levels of 3,500 sites/cell. Three distinct binding domains targeting CD22 were fused to various TCR signaling domains ± an IgG heavy chain constant domain (CH2CH3) to create a series of vector constructs suitable to delineate optimal CAR configuration. CARs derived from the m971 anti-CD22 moAb, which targets a proximal CD22 epitope demonstrated superior anti-leukemic activity compared to those incorporating other binding domains, and addition of a 4-1BB signaling domain to CD28.CD3ζ constructs diminished potency, whereas increasing affinity of the anti-CD22 binding motif, and extending the CD22 binding domain away from the membrane via CH2CH3 had no effect. We conclude that 2nd generation m971 derived anti-CD22-CARs are promising novel therapeutics that should be tested in BCP-ALL.
    Blood 12/2012; · 9.78 Impact Factor
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    ABSTRACT: We conducted a gene therapy trial in 10 patients with adenosine deaminase-deficient severe combined immunodeficiency (ADA-deficient SCID) using two slightly different retroviral vectors for the transduction of patients' bone marrow CD34+ cells. Four subjects were treated without pre-transplant cytoreduction and remained on ADA enzyme replacement therapy (ERT) throughout the procedure. Only transient (months), low level (<0.01%) gene marking was seen in peripheral blood mononuclear cells (PBMC) of two older subjects (15 and 20 years old), whereas some gene marking of PBMC has persisted for the past nine years in two younger subjects (4 and 6 years). Six additional subjects were treated using the same gene transfer protocol, but after withdrawal of ERT and administration of low-dose busulfan (65-90 mg/m(2)). Three of these remain well, off ERT (5, 4, and 3 years post-procedure), with gene marking in PBMC of 1-10%, and ADA enzyme expression in PBMC near or in the normal range. Two subjects were restarted on ERT due to poor gene marking and immune recovery and one had a subsequent allogeneic hematopoietic stem cell transplant. These studies directly demonstrate the importance of providing non-myeloablative pre-transplant conditioning to achieve therapeutic benefits with gene therapy for ADA-deficient SCID.
    Blood 09/2012; · 9.78 Impact Factor
  • Journal of Clinical Oncology 08/2012; · 18.04 Impact Factor
  • Alan S Wayne, Jerald P Radich
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    ABSTRACT: In the study reported in this issue of Blood by Leung et al, detectable minimal residual disease (MRD) before hematopoietic stem cell transplantation (HSCT) was found to be prognostic for outcome, but did not prevent cure for children with acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).
    Blood 07/2012; 120(2):244-6. · 9.78 Impact Factor

Publication Stats

911 Citations
348.97 Total Impact Points

Institutions

  • 2014
    • Children's Hospital Los Angeles
      • Children's Center for Cancer and Blood Diseases
      Los Angeles, California, United States
  • 2003–2014
    • National Institutes of Health
      • • Branch of Pediatric Oncology
      • • Center for Cancer Research
      Maryland, United States
  • 2004–2013
    • National Cancer Institute (USA)
      • • Center for Cancer Research
      • • Pediatric Oncology Branch
      Maryland, United States
  • 2012
    • Fred Hutchinson Cancer Research Center
      • Division of Clinical Research
      Seattle, Washington, United States
  • 2002–2011
    • NCI-Frederick
      Maryland, United States
  • 2010
    • University of Hamburg
      • Department of Stem Cell Transplantation
      Hamburg, Hamburg, Germany
  • 2009
    • National Cancer Institute
      Μπογκοτά, Bogota D.C., Colombia
  • 2008
    • George Washington University
      Washington, Washington, D.C., United States
  • 2006
    • National Institute of Mental Health (NIMH)
      Maryland, United States