Paclitaxel impairs adipose stem cell proliferation and differentiation

Abstract

BACKGROUND Cancer patients with chemotherapy-induced immunosuppression have poor surgical site wound healing. Prior literature supports the use of human adipose-derived stem cell (hASC) lipoinjection to improve wound healing. It has been established multipotent hASCs facilitate neovascularization, accelerated epithelialization, and wound closure in animal models. While hASC wound therapy may benefit surgical cancer patients, the chemotherapeutic effects on hASCs are unknown. We hypothesized Paclitaxel, a chemotherapeutic agent, impairs hASC growth, multipotency, and induces apoptosis. METHODS hASCs were isolated and harvested from consented, chemotherapy and radiation naïve patients. Growth curves, MTT, and EdU assays measured cytotoxicity and proliferation. Oil-Red-O stain, Alazarin-Red stain, Matrigel tube-formation assay, and qPCR analyzed hASC differentiation. Annexin V assay measured apoptosis. Immunostaining and Western blot determined TNF-α expression. RESULTS hASCs were selectively more sensitive to Paclitaxel (0.01μM–30μM) than fibroblasts (p<0.05). After 12 days, Paclitaxel caused hASC growth arrest whereas control hASCs proliferated (p=0.006). Paclitaxel caused an 80.6% reduction in new DNA synthesis (p<0.001). Paclitaxel severely inhibited endothelial differentiation and capillary-like tube formation. Differentiation markers LPL (adipogenic), alkaline phosphatase (osteogenic), CD31 and vWF (endothelial) were significantly decreased (all: p<0.05) confirming Paclitaxel impaired differentiation. Paclitaxel was also found to induce apoptosis and TNF-α was up-regulated in Paclitaxel-treated hASCs (p<0.001). CONCLUSION Paclitaxel is more cytotoxic to hASCs than fibroblasts. Paclitaxel inhibits hASC proliferation, differentiation, and induces apoptosis, possibly through the TNF-α pathway. Paclitaxel’s severe inhibition of endothelial differentiation indicates neovascularization disruption, possibly causing poor wound healing in cancer patients receiving chemotherapy.

Adipose-derived stem cell-mediated paclitaxel delivery inhibits breast cancer growth

“…On this point, some studies have been reported contradictory results; in fact, Huang et al [ 54 ] reported that human ASCs showed no cytotoxic effects when engulfed in nanoparticles containing even 30μM of PTX, instead others demonstrated that PTX-loaded ASCs, starting from 0.01μM, showed a reduced viability and growth [ 55 , 56 ]. However, most of the works have highlighted the harmlessness of PTX exposure (up to 1μM and beyond) on ASCs [ 39 , 43 , 54 , 57 ].…”

The current understanding of mesenchymal stem cells as potential attenuators of chemotherapy‐induced toxicity

“…The MSCs' proliferative ability seems strongly impaired by paclitaxel treatment, although the stem cells remained metabolically viable even after exposure to high doses . The level of apoptosis induction in paclitaxel‐treated MSCs has also been subject to controversy, with several datasets demonstrating slightly or strongly increased apoptosis, while only one publication reported no increase of apoptotic cells . On a molecular level, up‐regulation of TNF‐α after paclitaxel treatment was found associated with increased apoptosis in MSCs .…”

“…The level of apoptosis induction in paclitaxel‐treated MSCs has also been subject to controversy, with several datasets demonstrating slightly or strongly increased apoptosis, while only one publication reported no increase of apoptotic cells . On a molecular level, up‐regulation of TNF‐α after paclitaxel treatment was found associated with increased apoptosis in MSCs . Additionally, it has been demonstrated that paclitaxel resulted in a significant increase of premature senescence in MSCs, explaining the stem cells' observed preservation of viability despite a strong reduction in proliferation .…”

<p>The Role of Sharp Dissection in Nipple-Sparing Mastectomy: A Safe Procedure with No Necrosis of the Nipple-Areolar Complex</p>

“…These cytotoxic agents rapidly kill proliferating cells [31][32][33][34][35] but can impair regenerative cells, which are responsible for wound healing, and may increase the complication rates in the setting of breast reconstructive surgery. [35][36][37][38] However, based on our data, none of the patients with breast cancer showed poor wound healing, NAC discoloration, or ischemia. We demonstrated that NSM can be safe if offered to those who have received neoadjuvant chemotherapy or endocrine therapy.…”