Pilot Trial of FANG Immunotherapy in Ewing's Sarcoma

Maurizio Ghisoli, Minal Barve, Reva Schneider, Robert Mennel, Carl Lenarsky, Gladice Wallraven, Beena O Pappen, John LaNoue, Padmasini Kumar, Derek Nemunaitis, Alyssa Roth, James Nemunaitis, Sam Whiting, Neil Senzer, Frederick A Fletcher, John Nemunaitis, Maurizio Ghisoli, Minal Barve, Reva Schneider, Robert Mennel, Carl Lenarsky, Gladice Wallraven, Beena O Pappen, John LaNoue, Padmasini Kumar, Derek Nemunaitis, Alyssa Roth, James Nemunaitis, Sam Whiting, Neil Senzer, Frederick A Fletcher, John Nemunaitis

Abstract

We report on 12 consecutive patients with advanced/metastatic Ewing's sarcoma who were treated as a separate cohort of a phase 1 trial of FANG autologous immunotherapy (1 × 10(6)-2.5 × 10(7) cells/intradermal injection each month for minimum 4 months). Safety and clinical response were monitored. Patient immune response to unmodified autologous tumor cells was assessed by gamma interferon-enzyme-linked immunospot (γIFN-ELISPOT) assay using peripheral blood mononuclear cells from baseline (pretreatment) and multiple postvaccination time points. None of the 12 patients (47 vaccinations) developed grade 2/3/4 drug-related toxicity. Median product release granulocyte-macrophage colony-stimulating factor expression was 1,941 pg/10(6) cells, and TGFβ1and TGFβ2 knockdown were 99 and 100%, respectively. Eight patients were assessed for ELISPOT response to autologous tumor cells at baseline and all (100%) were negative. In contrast, follow-up ELISPOT response at month 1 or month 4 (one patient) after FANG was positive in all eight patients. One patient achieved a partial tumor response (38% tumor reduction, RECIST 1.1). The Kaplan-Meier estimated survival of these 12 patients at 1 year was 75%. In this phase 1 study in patients with Ewing's sarcoma, FANG immunotherapy was well tolerated, elicited a tumor-specific systemic immune response in all patients, and was associated with favorable 1-year survival. Further clinical testing is indicated.

Figures

Figure 1
Figure 1
Gamma interferon (γIFN) expression (enzyme-linked immunospot (ELISPOT)) of FANG vaccine-treated Ewing's sarcoma (EWS) patient PBMCs over time in response to nontransfected autologous tumor cells (n = 8). One patient had a second vaccine constructed with solitary lesion progression (patient #2, sample 098). Positive ELISPOT activation was again developed to the second harvested autologous tumor sample (green) (data as of 10/13/14). PBMC, peripheral blood mononuclear cell.
Figure 2
Figure 2
Kaplan–Meier survival curve of treated EWS patients (n = 12).
Figure 3
Figure 3
Status of patient #6 in Table 1: post frontline HD chemotherapy, vincristine/irinotcan/temodar, cixutumumab/timsirolimus, pazopanub/everolimus, ifosfamide/etoposide, meckinist/rapamycin/metformin, HD ifosfamide→surgery→FANG ×4.

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