PMID- 38040752 OWN - NLM STAT- MEDLINE DCOM- 20231206 LR - 20231221 IS - 2045-2322 (Electronic) IS - 2045-2322 (Linking) VI - 13 IP - 1 DP - 2023 Dec 1 TI - The Notch1 signaling pathway directly modulates the human RANKL-induced osteoclastogenesis. PG - 21199 LID - 10.1038/s41598-023-48615-2 [doi] LID - 21199 AB - Notch signaling is an evolutionary conserved pathway with a key role in tissue homeostasis, differentiation and proliferation. It was reported that Notch1 receptor negatively regulates mouse osteoclast development and formation by inhibiting the expression of macrophage colony-stimulating factor in mesenchymal cells. Nonetheless, the involvement of Notch1 pathway in the generation of human osteoclasts is still controversial. Here, we report that the constitutive activation of Notch1 signaling induced a differentiation block in human mononuclear CD14(+) cells directly isolated from peripheral blood mononuclear cells (PBMCs) upon in vitro stimulation to osteoclasts. Additionally, using a combined approach of single-cell RNA sequencing (scRNA-Seq) simultaneously with a panel of 31 oligo-conjugated antibodies against cell surface markers (AbSeq assay) as well as unsupervised learning methods, we detected four different cell stages of human RANKL-induced osteoclastogenesis after 5 days in which Notch1 signaling enforces the cell expansion of specific subsets. These cell populations were characterized by distinct gene expression and immunophenotypic profiles and active Notch1, JAK/STAT and WNT signaling pathways. Furthermore, cell-cell communication analyses revealed extrinsic modulators of osteoclast progenitors including the IL7/IL7R and WNT5a/RYK axes. Interestingly, we also report that Interleukin-7 receptor (IL7R) was a downstream effector of Notch1 pathway and that Notch1 and IL7R interplay promoted cell expansion of human RANKL-induced osteoclast progenitors. Taken together, these findings underline a novel cell pattern of human osteoclastogenesis, outlining the key role of Notch1 and IL-7R signaling pathways. CI - (c) 2023. The Author(s). FAU - Padovano, Costanzo AU - Padovano C AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Bianco, Salvatore Daniele AU - Bianco SD AD - Bioinformatics Unit, Fondazione IRCCS Casa Sollievo della Sofferenza, 71013, San Giovanni Rotondo, Italy. FAU - Sansico, Francesca AU - Sansico F AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - De Santis, Elisabetta AU - De Santis E AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Tamiro, Francesco AU - Tamiro F AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Colucci, Mattia AU - Colucci M AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Totti, Beatrice AU - Totti B AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Di Iasio, Serena AU - Di Iasio S AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Bruno, Gaja AU - Bruno G AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Panelli, Patrizio AU - Panelli P AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Miscio, Giuseppe AU - Miscio G AD - Clinical Laboratory Analysis and Transfusional Medicine, Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. FAU - Mazza, Tommaso AU - Mazza T AD - Bioinformatics Unit, Fondazione IRCCS Casa Sollievo della Sofferenza, 71013, San Giovanni Rotondo, Italy. FAU - Giambra, Vincenzo AU - Giambra V AD - Hematopathology Laboratory, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS "Casa Sollievo della Sofferenza", 71013, San Giovanni Rotondo (FG), Italy. v.giambra@operapadrepio.it. LA - eng GR - GR-2016-02361287/Ministero della Salute/ PT - Journal Article DEP - 20231201 PL - England TA - Sci Rep JT - Scientific reports JID - 101563288 RN - 0 (RANK Ligand) RN - 0 (TNFSF11 protein, human) RN - 0 (NOTCH1 protein, human) SB - IM MH - Humans MH - Cell Differentiation MH - *Leukocytes, Mononuclear MH - Osteoclasts/metabolism MH - *Osteogenesis MH - RANK Ligand/pharmacology/metabolism MH - Signal Transduction PMC - PMC10692129 COIS- The authors declare no competing interests. EDAT- 2023/12/02 00:42 MHDA- 2023/12/04 12:42 PMCR- 2023/12/01 CRDT- 2023/12/01 23:25 PHST- 2023/06/08 00:00 [received] PHST- 2023/11/28 00:00 [accepted] PHST- 2023/12/04 12:42 [medline] PHST- 2023/12/02 00:42 [pubmed] PHST- 2023/12/01 23:25 [entrez] PHST- 2023/12/01 00:00 [pmc-release] AID - 10.1038/s41598-023-48615-2 [pii] AID - 48615 [pii] AID - 10.1038/s41598-023-48615-2 [doi] PST - epublish SO - Sci Rep. 2023 Dec 1;13(1):21199. doi: 10.1038/s41598-023-48615-2.