Thalidomide induces apoptosis during early mesodermal differentiation of human induced pluripotent stem cells

Abstract

Thalidomide was once administered to pregnant women as a mild sedative; however, it was subsequently shown to be strongly teratogenic. Recently, there has been renewed interest in thalidomide because of its curative effects against intractable diseases. However, the teratogenicity of thalidomide is manifested in various ways and is still not fully understood. In the present study, we evaluated the effects of thalidomide on early mesodermal differentiation by examining the differentiation of human induced pluripotent stem cells (hiPSCs). The most common symptom of thalidomide teratogenicity is limb abnormality, which led us to hypothesize that thalidomide prevents early mesodermal differentiation. Therefore, mesodermal differentiation of hiPSCs was induced over a 6-d period. To induce early mesoderm differentiation, 1 d after seeding, the cells were incubated with the small molecule compound CHIR99021 for 3 d. Thalidomide exposure was initiated at the same time as CHIR99021 treatment. After 5 d of thalidomide exposure, the hiPSCs began expressing a mesodermal marker; however, the number of viable cells decreased significantly as compared to that of control cells. We observed that the proportion of apoptotic and dead cells increased on day 2; however, the proportion of dead cells on day 5 had decreased, suggesting that the cells were damaged by thalidomide during early mesodermal differentiation (days 0-2). Our findings may help elucidate the mechanism underlying thalidomide teratogenicity and bring us closer to the safe use of this drug.

Intrauterine exposure to drugs and reproduction—still reasons for concern!

“…As thalidomide has multiple effects on body homeostasis, the specific mechanisms remain debated. It has been suggested that thalidomide disrupts the early mesodermal differentiation by damaging undifferentiated cells (Tachikawa et al 2018) as well as resulting in anti-angiogenesis, evasion of apoptosis and inhibition of pro-inflammatory innate responses (Paravar and Lee 2008;Hideshima et al 2000;Du et al 2005). Furthermore, thalidomide can bind as an antagonist to the androgen receptor (AR) (Capitosti et al 2004) and hence inhibit different androgens like testosterone which have a pivotal impact during development of male secondary sex characteristics and reproductive tissues.…”

Thalidomide Inhibits Human iPSC Mesendoderm Differentiation by Modulating CRBN-dependent Degradation of SALL4

“…Human pluripotent stem cells have proven useful in investigating the teratogenicity of thalidomide in vitro. Studies on the effect of thalidomide on human pluripotent stem cells (hPSCs) characterized cytotoxicity in undifferentiated hPSCs 19,20 and hPSC-derived mesoderm 21 and functional effects on hPSC-derived mesendoderm 22 and cardiac mesoderm 20 at concentrations above 100 µM. However, these studies demonstrated effects of thalidomide on PSCs or PSC-derived mesoderm in vitro at concentrations >5-fold in excess of the thalidomide clinical Cmax 7 , which raises concern regarding the relevance of these effects to thalidomide teratogenicity.…”