Effect of Collagen Matrix on Doxorubicin Distribution and Cancer Cells’ Response to Treatment in 3D Tumor Model

Abstract

The extracellular matrix (ECM) plays an important role in regulation of many aspects of tumor growth and response to therapies. However, the specifics of the interaction of chemotherapeutic agents with cancer cells in the presence of collagen, the major component of ECM, is still poorly investigated. In this study, we explored distribution of doxorubicin (DOX) and its effects on cancer cells’ metabolism in the presence of collagen with different structures in 3D models. For this, a combination of second harmonic generation imaging of collagen and multiphoton fluorescence microscopy of DOX, and metabolic cofactor NAD(P)H was used. It was found that collagen slowed down the diffusion of DOX and thus decreased the cellular drug uptake. Besides nuclei, DOX also targeted mitochondria leading to inhibition of oxidative phosphorylation, which was more pronounced in the cells growing in the absence of collagen. As a result, the cells in collagen displayed better viability upon treatment with DOX. Taken together, our data illustrate that tumor collagen contributes to heterogeneous and sub-optimal response to DOX and highlight the challenges in improving drug delivery and efficacy.

Monitoring the Intracellular pH and Metabolic State of Cancer Cells in Response to Chemotherapy Using a Combination of Phosphorescence Lifetime Imaging Microscopy and Fluorescence Lifetime Imaging Microscopy

“…It was noticed that, upon long-term (>48 h) cultivation in collagen, the HCT116 cells showed a tendency toward a decrease in the NAD(P)H τ mean and an increase in the a 1 /a 2 ratio, which could be associated with either enhanced glycolysis or reduced mitochondrial respiration ( Figure 4 ). The latter is more likely, because cells growing in collagen have lower proliferation and metabolic rates [ 38 ].…”

“…Therefore, it is not surprising that the FOLFOX combination of drugs produced the same effect. There are several other works that demonstrate an induction of mitochondrial OXPHOS activity in response to chemotherapy by using the FLIM of NAD(P)H [ 24 , 38 ].…”

“…As for the effects of the tumor extracellular matrix on the efficacy of chemotherapy, the obtained results clearly indicate that the presence of unstructured collagen reduces the therapeutic efficacy of the FOLFOX regimen against colon cancer cells, as assessed from the cell viability, pHi and metabolic assays. The influence of the ECM on tumor responses to chemotherapy is well documented [ 38 , 52 , 53 , 54 , 55 , 56 , 57 , 58 ]. In our recent study on T24 (human urinary bladder carcinoma) cells growing in 3D collagen-based models, we showed that collagen protects cancer cells from the action of doxorubicin, improves viability and diminishes cellular metabolic changes upon treatment [ 38 ].…”

“…The influence of the ECM on tumor responses to chemotherapy is well documented [ 38 , 52 , 53 , 54 , 55 , 56 , 57 , 58 ]. In our recent study on T24 (human urinary bladder carcinoma) cells growing in 3D collagen-based models, we showed that collagen protects cancer cells from the action of doxorubicin, improves viability and diminishes cellular metabolic changes upon treatment [ 38 ]. In the paper by Chen et al, a low TACS (Tumor-Associated Collagen Signature, the index that positively correlates with collagen area, collagen straightness and collagen cross-link density) level in gastric tumors was associated with improved outcomes in patients who received 5-fluorouracil-based adjuvant chemotherapy [ 55 ].…”

New Methacrylated Biopolymer-Based Hydrogels as Localized Drug Delivery Systems in Skin Cancer Therapy

“…Meanwhile, the influence of the drug on the cell division process can be observed in the fluorescence microscopy images as empty areas on the culture plate well, as compared to non-exposed cells. Further studies will be performed in order to elucidate the effect of Dox in A431 tumoral cells (with biomarkers such as AMPK, p53, and Bcl-2, which have been identified as important for apoptosis induction by doxorubicin [64]) and to correlate the concentration of the drug loaded in the hydrogels with its biological effects.…”