Highly Invasive Fluorescent/Bioluminescent Patient-Derived Orthotopic Model of Glioblastoma in Mice

Abstract

Development of the novel diagnostic and therapeutic approaches in neuro-oncology requires tumor models that closely reproduce the biological features of patients’ tumors. Patient-derived xenografts (PDXs) are recognized as a valuable and the most “close-to-patient” tool for preclinical studies. However, their establishment is complicated by the factors related to both the surgical material and technique of the orthotopic implantation. The aim of this work was to develop a patient-derived glioblastoma multiform (GBM) model that stably co-expresses luciferase and a far-red fluorescent protein for monitoring of tumor progression in the brain and, using this model, to validate new diagnostic methods—macroscopic fluorescence lifetime imaging (macro-FLIM) and cross-polarization optical coherence tomography (CP OCT). The established model was similar to the original patient’s GBM in terms of histological and immunohistochemical features and possessed reproducible growth in nude mice, which could be observed by both fluorescence and bioluminescence imaging. Our results demonstrated the high potential of macro-FLIM and CP OCT for intraoperative differentiation of GBM from the white matter. Thus, the dual-labeled PDX model of GBM proved to be an excellent approach for observation of tumor development by optical methods.

Breast Cancer Brain Metastases: Implementation and Characterization of a Mouse Model Relying on Malignant Cells Inoculation in the Carotid Artery

“…Alternative injection routes primarily targeting the brain, such as direct intracerebral, arose to induce experimental BCBM through the puncturing of the mice’s skull [ 20 , 21 ]. Despite its utility in leptomeningeal melanoma metastases research [ 22 ], direct intracerebral injection is most promising in primary local growth models, such as glioblastoma [ 23 ], rather than metastatic ones, as it can only replicate the final stage of the metastatic process [ 14 ]. Importantly, the impact of the administration route on therapeutics’ uptake [ 24 ], where the intracranial injection may induce alterations on the blood–brain barrier (BBB) and blood–tumour barrier, has been demonstrated, increasing the brain penetration of non-targeted pharmaceuticals associated with the injection method, which does not occur in intracardiac and tail intravenous injection routes [ 24 ].…”

“…Although in vivo models have been put into place to effectively deliver cells into the brain [ 23 ] and promote preferential BM formation [ 16 , 20 ], due to the nature of certain procedures, these models may lead to unfortunate outcomes prior to BM development. Therefore, the intracarotid injection represents a preferential highway to the brain, allowing cancer cells’ efficient delivery to this site.…”

Phototoxicity and cell passage affect intracellular reactive oxygen species levels and sensitivity towards non-thermal plasma treatment in fluorescently-labeled cancer cells

“…These considerations should not only be taken into account for experiments utilizing 2D monolayer cultures demonstrated here, but also for those using 3D in vitro models. Moreover, immunofluorescence is also used in in vivo research for tracking tumor growth in orthotopic models and follow-up of metastasis [43,44]. Although ambient light is not a confounding factor in situ in the in vivo environment, our findings could have important implications to take into account during preparatory cell culturing and experimental handling prior to tumor cell inoculation.…”

Metabolic heterogeneity of colorectal cancer as a prognostic factor: insights gained from fluorescence lifetime imaging

“…Recently, we demonstrated a high variability of cellular metabolic statuses in CT26 tumors of large size compared with small tumors, which correlated with heterogeneous oxygen distribution (Parshina et al, 2022). A significant intertumor heterogeneity of metabolism was observed in patient-derived glioblastoma xenografts, which differed them from standard U87 glioma (Yuzhakova et al, 2022).…”

“…For example, tumor spheroids obtained from the cervical cancer cell line HeLa (Lukina et al, 2018) or murine colorectal carcinoma cell line CT26 (Shirmanova et al, 2021) displayed the differences in metabolism between the outer and the inner cell layers with the outer layers being more glycolytic (higher NAD(P)H-a 1 ). The spheroids generated from the patients' derived glioblastoma cultures did not show metabolic zonality, but generally had greater intercellular variations of NAD(P)H lifetime parameters than the spheroids from standard line U373 MG (Yuzhakova et al, 2023). A spectrum of works by M.…”