iPSC-derived lung and lung cancer organoid model to evaluate cisplatin encapsulated autologous iPSC-derived mesenchymal stromal cell-isolated extracellular vesicles

Abstract

Background Lung cancer remains a leading cause of cancer-related mortality globally. Although recent therapeutic advancements have provided targeted treatment approaches, the development of resistance and systemic toxicity remain primary concerns. Extracellular vesicles (EVs), especially those derived from mesenchymal stromal cells (MSC), have gained attention as promising drug delivery systems, offering biocompatibility and minimal immune responses. Recognizing the limitations of conventional 2D cell culture systems in mimicking the tumor microenvironment, this study aims to describe a proof-of-principle approach for using patient-specific organoid models for both lung cancer and normal lung tissue and the feasibility of employing autologous EVs derived from induced pluripotent stem cell (iPSC)-MSC in personalized medicine approaches. Methods First, we reprogrammed healthy fibroblasts into iPSC. Next, we differentiated patient-derived iPSC into branching lung organoids (BLO) and generated patient-matched lung cancer organoids (LCO) from patient-derived tumor tissue. We show a streamlined process of MSC differentiation from iPSC and EV isolation from iPSC-MSC, encapsulated with 0.07 µg/mL of cytotoxic agent cisplatin and applied to both organoid models. Cytotoxicity of cisplatin and cisplatin-loaded EVs was recorded with LDH and CCK8 tests. Results Fibroblast-derived iPSC showed a normal karyotype, pluripotency staining, and trilineage differentiation. iPSC-derived BLO showed expression of lung markers, like TMPRSS2 and MUC5A while patient-matched LCO showed expression of Napsin and CK5. Next, we compared the effects of iPSC-MSC derived EVs loaded with cisplatin against empty EVs and cisplatin alone in lung cancer organoid and healthy lung organoid models. As expected, we found a cytotoxic effect when LCO were treated with 20 µg/mL cisplatin. Treatment of LCO and BLO with empty EVs resulted in a cytotoxic effect after 24 h. However, EVs loaded with 0.07 µg/mL cisplatin failed to induce any cytotoxic effect in both organoid models. Conclusion We report on a proof-of-principle pipeline towards using autologous or allogeneic iPSC-MSC EVs as drug delivery tests for lung cancer in future. However, due to the time and labor-intensive processes, we conclude that this pipeline might not be feasible for personalized approaches at the moment.

1. Introduction

The World Health Organization (WHO) predicts that, within the next forty years, cancer will surpass ischemic heart disease (IHD) as the primary cause of death, demonstrating a projected increase of more than two-fold from 2016 to 2060 [1]. With 2.2 million new cases and 1.8 million deaths worldwide in 2020, lung cancer is one of the most common types of cancer and a leading cause of cancer-related deaths worldwide [2]. Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer cases [3].

In the context of lung cancer treatment, while recent breakthroughs in targeted molecular therapy and immuno-oncology have transformed the field, cytotoxic chemotherapy remains the primary therapeutic option [4]. Traditional systemic administration of chemotherapy drugs, such as cisplatin, often leads to significant toxicity due to the lack of specificity of cancer cells [5]. In recent years, targeted approaches have emerged, particularly in cases with driver mutations such as EGFR exon 19 deletion or ALK gene translocation [6]. However, it is important to note that almost all targeted therapy approaches eventually develop resistance, rendering the therapy ineffective [7]. Furthermore, targeted therapy is only suitable for a small percentage of patients, as not all cases exhibit predominant driver mutations [8]. Hence, an urgent requirement exists for drug delivery systems with the capacity to transport drugs directly to tumors, thus mitigating systemic toxicity. Innovative strategies, such as nanoparticle-based delivery systems, hold great promise for addressing this need.