Front Oncol 9: 1034

PMC: PMC6823179

Dendritic Cell-Based Immunotherapy in Advanced Sarcoma and Neuroblastoma Pediatric Patients: Anti-cancer Treatment Preceding Monocyte Harvest Impairs the Immunostimulatory and Antigen-Presenting Behavior of DCs and Manufacturing Process Outcome

Lucie Jureckova+8 authors

Supplementary Figure 1

Decision tree for DC-IMP manufacturing workflow including in-process and quality controls.

Click here for additional data file.^{(268K, pdf)}

Supplementary Figure 2

Number of anti-cancer therapy lines preceding monocyte harvest were compared based on QC parameters: (A) DC yield, and post-thaw: (B) viability, (C) DC phenotype on day 0: CD14, CD197, CD80, CD86, CD83 and on day 2: MHC II, CD80, CD86, CD83 and immunostimulatory properties presented by (D) IL-12 production, IL-10 production and IL-12/IL-10 production ratio, (E) allo-MLR and auto-MLR.

Click here for additional data file.^{(842K, tif)}

Supplementary Figure 3

Manufacturing subgroup from monocytes harvested after MTD-based therapy potentially interfering with monocyte biology and manufacturing subgroup from monocytes from untreated patients or after non-interfering treatment compared based on QC parameters: (A) DC yield, and post-thaw: (B) viability, (C) DC phenotype on day 0: CD14, CD197, CD80, CD86, CD83 and on day 2: MHC II, CD80, CD86, CD83 and immunostimulatory properties presented by (D) IL-12 production, IL-10 production and IL-12/IL-10 production ratio, (E) allo-MLR and auto-MLR.

Click here for additional data file.^{(857K, tif)}

Supplementary Table 1

Monocyte biology-interfering medications.

Click here for additional data file.^{(65K, DOCX)}

Supplementary Table 2

Study patient characteristics, disease course, and therapy.

Click here for additional data file.^{(85K, DOCX)}

Supplementary Table 3

Source data: CBC parameters, manufacturing details, and QC parameters.

Click here for additional data file.^{(19K, XLSX)}

Supplementary Material 1

html. Interactive—medications 60 days prior to monocyte harvest.

Click here for additional data file.^{(964K, ZIP)}

^{Department of Pharmacology, Faculty of Medicine, Masaryk University, Brno, Czechia}

^{Department of Pediatric Oncology, University Hospital and Faculty of Medicine, Masaryk University, Brno, Czechia}

^{Regional Centre for Applied Molecular Oncology, Masaryk Memorial Cancer Institute, Brno, Czechia}

^{Department of Internal Medicine-Hematology and Oncology, University Hospital and Medical Faculty, Masaryk University, Brno, Czechia}

^{Transfusion and Tissue Department, University Hospital Brno, Brno, Czechia}

Edited by: Christina Annunziata, National Cancer Institute (NCI), United States

Reviewed by: Daniel Green, Kite Pharma, United States; Oladapo Yeku, Massachusetts General Hospital Cancer Center, United States

*Correspondence: Lenka Zdrazilova-Dubska zc.uom@aksbud

This article was submitted to Cancer Molecular Targets and Therapeutics, a section of the journal Frontiers in Oncology

†These authors have contributed equally to this work and are listed in alphabetical order

Edited by: Christina Annunziata, National Cancer Institute (NCI), United States

Reviewed by: Daniel Green, Kite Pharma, United States; Oladapo Yeku, Massachusetts General Hospital Cancer Center, United States

Received 2019 Jul 19; Accepted 2019 Sep 24.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

Abstract

Despite efforts to develop novel treatment strategies, refractory and relapsing sarcoma, and high-risk neuroblastoma continue to have poor prognoses and limited overall survival. Monocyte-derived dendritic cell (DC)-based anti-cancer immunotherapy represents a promising treatment modality in these neoplasias. A DC-based anti-cancer vaccine was evaluated for safety in an academic phase-I/II clinical trial for children, adolescents, and young adults with progressive, recurrent, or primarily metastatic high-risk tumors, mainly sarcomas and neuroblastomas. The DC vaccine was loaded with self-tumor antigens obtained from patient tumor tissue. DC vaccine quality was assessed in terms of DC yield, viability, immunophenotype, production of IL-12 and IL-10, and stimulation of allogenic donor T-cells and autologous T-cells in allo-MLR and auto-MLR, respectively. Here, we show that the outcome of the manufacture of DC-based vaccine is highly variable in terms of both DC yield and DC immunostimulatory properties. In 30% of cases, manufacturing resulted in a product that failed to meet medicinal product specifications and therefore was not released for administration to a patient. Focusing on the isolation of monocytes and the pharmacotherapy preceding monocyte harvest, we show that isolation of monocytes by elutriation is not superior to adherence on plastic in terms of DC yield, viability, or immunostimulatory capacity. Trial patients having undergone monocyte-interfering pharmacotherapy prior to monocyte harvest was associated with an impaired DC-based immunotherapy product outcome. Certain combinations of anti-cancer treatment resulted in a similar pattern of inadequate DC parameters, namely, a combination of temozolomide with irinotecan was associated with DCs showing poor maturation and decreased immunostimulatory features, and a combination of pazopanib, topotecan, and MTD-based cyclophosphamide was associated with poor monocyte differentiation and decreased DC immunostimulatory parameters. Searching for a surrogate marker predicting an adverse outcome of DC manufacture in the peripheral blood complete blood count prior to monocyte harvest, we observed an association between an increased number of immature granulocytes in peripheral blood and decreased potency of the DC-based product as quantified by allo-MLR. We conclude that the DC-manufacturing yield and the immunostimulatory quality of anti-cancer DC-based vaccines generated from the monocytes of patients were not influenced by the monocyte isolation modality but were detrimentally affected by the specific combination of anti-cancer agents used prior to monocyte harvest.

Keywords: dendritic cells, anti-cancer medications, sarcoma, neuroblastoma, cell-based medicinal products, investigator-initiated clinical trial, manufacturing outcome variability

Abstract

Footnotes

Funding. This work was supported by the Czech Ministry of Education, Youth and Sport via Large infrastructure CZECRIN (LM2015090) and RECAMO (LO1413), by the European Regional Development Fund—project CZECRIN_4PATIENTY (Reg. No. CZ.02.1.01/0.0/0.0/16_013/0001826), and by the Czech Ministry of Health via DRO 00209805.

Footnotes

Click here for additional data file.^{(268K, pdf)}Click here for additional data file.^{(842K, tif)}Click here for additional data file.^{(857K, tif)}Click here for additional data file.^{(65K, DOCX)}Click here for additional data file.^{(85K, DOCX)}Click here for additional data file.^{(19K, XLSX)}Click here for additional data file.^{(964K, ZIP)}

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