Development of next-generation tumor-homing induced neural stem cells to enhance treatment of metastatic cancers

Abstract

Engineered tumor-homing neural stem cells (NSCs) have shown promise in treating cancer. Recently, we transdifferentiated skin fibroblasts into human-induced NSCs (hiNSC) as personalized NSC drug carriers. Here, using a SOX2 and spheroidal culture-based reprogramming strategy, we generated a new hiNSC variant, hiNeuroS, that was genetically distinct from fibroblasts and first-generation hiNSCs and had significantly enhanced tumor-homing and antitumor properties. In vitro, hiNeuroSs demonstrated superior migration to human triple-negative breast cancer (TNBC) cells and in vivo rapidly homed to TNBC tumor foci following intracerebroventricular (ICV) infusion. In TNBC parenchymal metastasis models, ICV infusion of hiNeuroSs secreting the proapoptotic agent TRAIL (hiNeuroS-TRAIL) significantly reduced tumor burden and extended median survival. In models of TNBC leptomeningeal carcinomatosis, ICV dosing of hiNeuroS-TRAIL therapy significantly delayed the onset of tumor formation and extended survival when administered as a prophylactic treatment, as well as reduced tumor volume while prolonging survival when delivered as established tumor therapy.

Use of FLOSEAL® as a scaffold and its impact on induced neural stem cell phenotype, persistence, and efficacy

“…“Day 0 iNSCs” denotes fibroblasts that have been transduced and transdifferentiated to become iNSCs but not placed into a scaffold. NSC, differentiation, proliferation, pluripotency, migration, therapy, and anti‐apoptosis markers were monitored; specific genes were selected based on bulk and single‐cell RNA sequencing conducted previously by our group 26,27 . The differentiation markers, namely GFAP , TUBB3 , and VMAC , were found to remain fairly constant in their expression levels over time, but downregulated compared to the day 0 iNSCs (Figure 2b).…”

“…NSC, differentiation, proliferation, pluripotency, migration, therapy, and anti-apoptosis markers were monitored; specific genes were selected based on bulk and single-cell RNA sequencing conducted previously by our group. 26,27 The differentiation markers, namely GFAP, TUBB3, and VMAC, were found to remain fairly constant in their expression levels over time, but downregulated compared to the day 0 iNSCs (Figure 2b). Of the NSC markers, NESTIN was the only one found to be upregulated (Figure 2c).…”

Utilizing induced neural stem cell‐based delivery of a cytokine cocktail to enhance chimeric antigen receptor‐modified T‐cell therapy for brain cancer

“…An IRES element driving green fluorescent protein (GFP) expression was also included to quantify transduction efficiency. Using our well‐established methodology for converting human fibroblasts into second‐generation iNSCs, human‐induced neurospheres (hiNeuroS), 17 we engineered RANTES/IL‐15‐secreting hiNeuroS (hiNeuroS RANTES‐IL‐15 ) (Figure 2b). Robust expression of the GFP reporter gene was detected by fluorescence microscopy (Figure 2c).…”

Spatiotemporal analysis of induced neural stem cell therapy to overcome advanced glioblastoma recurrence

“… 29 We speculate that the inconsistent results obtained from using ICV therapy in these mice may be due to mouse-to-mouse variability in ventricular displacement. Our group has previously modeled ICV injection of therapeutic cells, 30 , 31 but not in a model of large, late-stage brain cancer. Further use of ICV infusion against this tumor model may require correlative imaging techniques as seen in the clinic.…”