Phthalimide Derivative Shows Anti-angiogenic Activity in a 3D Microfluidic Model and No Teratogenicity in Zebrafish Embryos

Abstract

Angiogenesis is a crucial event for tumor progression and metastasis. It is the process through which new blood vessels are formed and has become a therapeutic target in many cancer therapies. However, current anti-angiogenic drugs such as Thalidomide still have detrimental teratogenic effects. This property could be caused by the presence of chiral carbons, intrinsic to such compounds. We synthesized four different phthalimide derivatives that lack chiral carbons in their chemical structure. We hypothesized that these achiral carbon compounds would retain similar levels of anti-angiogenic activity whilst reducing teratogenic effects. We tested for their anti-angiogenic functions using an in vitro 3D microfluidic assay with human endothelial cells. All four compounds caused a drastic inhibition of angiogenesis at lower effective concentrations compared to Thalidomide. Quantification of the blood vessel sprouting in each condition allowed us to classify compounds depending on their anti-angiogenic capabilities. The most effective identified compound (C4), was tested in vivo on a zebrafish embryo model. Blood vessel development was measured using number and lengths of the stalks visible in the fli1a :EGFP transgenic line. Potential teratogenic effects of C4 were monitored over zebrafish embryonic development. The in vivo results confirmed the increased potency of C4 compared to Thalidomide demonstrated by results in embryos exposed to concentrations as low as 0.02 μM. The teratogenic analysis further validated the advantages of using C4 over Thalidomide in zebrafish embryos. This study highlights how the use of in vitro 3D model can allow rapid screening and selection of new and safer drugs.

A 3D pancreatic tumor model to study T cell infiltration

“…[25][26][27] 3D microfluidic models allow a real-time analysis of cellular interactions, the possibility to mimic the main steps of the metastatic cascade and elucidate critical factors in the tumor progression. [28][29][30][31][32][33] For instance, the involvement of macrophages in regulating the intravasation of breast cancer cells through an endothelial layer when co-cultured in a microfluidic device was demonstrated by Zervantonakis et al 34 In a microfluidic-based lung carcinoma model, Bai et al investigated the contribution of tumor-associated macrophages (TAMs) in modulating the epithelial-to-mesenchymal transition (EMT) of cancer cell aggregates. 35 Additionally, Penny et al showed that TAMs promote PDAC cell extravasation through a vascular wall by co-culturing cancer cells, macrophages and endothelial cells (ECs) in a microfluidic device.…”

Evaluating Nanoparticles in Preclinical Research Using Microfluidic Systems

“…Currently microfluidic systems have been used to culture various small organisms such as zebrafish [139,140], C. elegans [141,142] and fruit flies [143]. Despite that some of these models have been applied for chemical evaluations [144,145,146], few have been adopted in NP evaluation. C. elegans is one of the model organisms which have been applied for NP testing.…”

3D tumor angiogenesis models: recent advances and challenges

“…Many studies have attempted to mimic the angiogenic tumor microenvironment using microfluidic devices. One such device was used to show the role of phthalimide compounds in reducing angiogenesis by observing the sprouting ability of endothelial cells after treatment (Mercurio et al 2019 ). Treatment of tumor organoids with tyrosine kinase inhibitor sorafenib showed a disturbance in the formation of endothelial networks.…”