Single-Cell RNA Sequencing of Human Embryonic Stem Cell Differentiation Delineates Adverse Effects of Nicotine on Embryonic Development

Abstract

Summary Nicotine, the main chemical constituent of tobacco, is highly detrimental to the developing fetus by increasing the risk of gestational complications and organ disorders. The effects of nicotine on human embryonic development and related mechanisms, however, remain poorly understood. Here, we performed single-cell RNA sequencing (scRNA-seq) of human embryonic stem cell (hESC)-derived embryoid body (EB) in the presence or absence of nicotine. Nicotine-induced lineage-specific responses and dysregulated cell-to-cell communication in EBs, shedding light on the adverse effects of nicotine on human embryonic development. In addition, nicotine reduced cell viability, increased reactive oxygen species (ROS), and altered cell cycling in EBs. Abnormal Ca 2+ signaling was found in muscle cells upon nicotine exposure, as verified in hESC-derived cardiomyocytes. Consequently, our scRNA-seq data suggest direct adverse effects of nicotine on hESC differentiation at the single-cell level and offer a new method for evaluating drug and environmental toxicity on human embryonic development in utero .

Effects of cigarette smoke components on myocardial differentiation of mouse embryonic stem cells

“…A previous study reported that nicotine exposure did not affect cell survival in mESCs when applied at 0.1‐10 μM for 48 hours, but that cigarette smoke condensate (CSC) changed cell proliferation and cell doubling time and induced cell death . Other previous studies showed a slight decrease in response to exposure to 10 μM nicotine for 21 days, but this difference was not great when compared to the control group; rather, the cell cycle of the EBs was changed . These previous studies provide evidence that nicotine showed a change in cell cycle‐related proteins such as p53 and p21 (but not HDAC1), and did not significantly affect the size of mEB.…”

“…Cigarette smoke contains over 4000 chemicals, and a significant number of substances and their metabolites pass through the placenta, affecting embryonic, and neonatal development of major organs, including heart . Nicotine, a major chemical component of cigarettes, is also extremely harmful to fetal development by increasing the risk of pregnancy complications and long‐term disease . Therefore, maternal smoking during pregnancy may be one of the causative factors for low birth weight and common diseases, including cardiovascular disease and obesity, in adulthood .…”

Latent periodic process inference from single-cell RNA-seq data

“…Broader applications of Cyclum. To evaluate Cyclum on larger high-throughput single-cell RNA-seq dataset, we applied it on a human embryonic stem cell (hESC) dataset containing 12,280 cells generated by the 10X Chromium technology, from a study of nicotine effects 38 . Results show that Cyclum accurately and efficiently infers cell-cycle pseudo-time ( Supplementary Fig.…”

“…8a and Supplementary Note 2). Targets of nicotine, including LDHA 38 and ENO1 39 are proposed by evaluating the difference of circular pattern in each gene between the treated sample and the control sample ( Supplementary Fig. 8b and Supplementary Note 2).…”

Maternal Smoking Induced Cardiovascular Risks in Fetuses: How Can in silico Models Help?

“…In particular, the human induced pluripotent stem cells (hiPSCs) can be derived from patient-and ethnic-specific background and hence can reproduce different kinds of cell types of human organs. For example, the change in Ca 2+ homeostasis due to nicotine exposure was recently demonstrated by using human embryonic stem cells (hESCs; Guo et al, 2019). The same group reported that e-cigarettes caused dysfunction of hiPSC-derived ECs .…”