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The paracrine interaction between tumor cells and adjacent stroma has been associated with the oncogenic activity of the Hedgehog (Hh) pathway in triple-negative breast tumors. The present study developed a model of paracrine Hh signaling and examined the impact of mesenchymal cell sources and culture modalities in the oncogenicity of the Hh pathway in breast tumor cells. Studies consisted of tumor cell monocultures and co-cultures with cancer-associated and normal fibroblasts, tumor cells that undergo epithelial–mesenchymal transition (EMT), or adipose-derived mesenchymal stem cells (ADMSCs). Hh ligand and pathway inhibitors, GANT61 and NVP-LDE225 (NVP), were evaluated in both cell cultures and a mouse xenograft model. Results in monocultures show that tumor cell viability and Hh transcriptional activity were not affected by Hh inhibitors. In co-cultures, down-regulation of GLI1, SMO, and PTCH1 in the stroma correlated with reduced tumor growth rates in xenografted tumors and cell cultures, confirming a paracrine interaction. Fibroblasts and EMT cells supported Hh transcriptional activity and enhanced tumor cell growth. Mixed and adjacent culture modalities indicate that tumor growth is supported via fibroblast-secreted soluble factors, whereas enriched tumor stemness requires close proximity between tumor and fibroblasts. Overall this study provides a tumor–mesenchymal model of Hh signaling and highlights the therapeutic value of mesenchymal cells in the oncogenic activity of the Hh pathway. [ABSTRACT FROM AUTHOR]

Gene editing is becoming the plant breeding tool of choice, but prior to targeting a gene for editing, a knowledge of the gene family members (GFMs) controlling yield is required in the specific crop plant. Critical to yield are components from senescing leaves. We targeted genes controlling senescence in Pisum sativum and the release and transport of carbohydrates and amino acids from the source leaves to the pods and seeds. The expression of GFMs for cytokinin biosynthesis (IPT) and destruction (CKX), sucrose transporters (SUT), Sugar Will Eventually be Exported Transporters (SWEET), amino acid permeases (AAP), and cell wall invertases, was determined using RT-qPCR. GFMs were differentially expressed in leaves of different ages. The expression of many gene family members was lower in the expanding sink leaves compared with the senescing leaves, with the exception of two PsAAP GFMs and PsCKX5, which were highly expressed. Expression of specific PsSWEETs, SUTs, and AAPs increased in the mature and/or senescing leaves. Expression of PsIPTs was least in the mature source leaves, but as strong in the senescing leaves as in the young source leaves. PsCKX7 was expressed in source and senescing leaves. We discuss the potential impact of the targeted reduction of specific PsCKX GFMs on source-sink relationships. [ABSTRACT FROM AUTHOR]