Growth Factors and Signaling in Development and Disease: (last updated 08/02/22)

The major focus of my laboratory has been to identify colony stimulating factor-1 (CSF-1), its 3 isoforms and its receptor (CSF-1R), and to define their developmental and physiological roles using biochemical, cell biological and mouse genetic approaches. We have shown that CSF-1 and the CSF-1R regulate the production and maintenance of macrophages, osteoclasts, microglia, Langerhans cells, Paneth cells and neural progenitor cells and play an important role in the development of leukemia and of several inflammatory diseases. We have shown that CSF-1 and the CSF-1R, via their regulation of tumor-associated macrophages, enhance solid tumor progression. We have developed novel biochemical and genetic approaches to CSF-1R signal transduction to analyze CSF-1R structure/function, identifying and elucidating the function of several downstream signaling molecules, including protein tyrosine phosphatase SHP-1, that negatively affects cell survival in the absence of CSF-1, the cbl proto-oncogene product, that negatively regulates CSF-1 proliferation signaling by enhancing CSF-1R endocytosis and the protein tyrosine phosphatase, PTP-phi, that mediates CSF-1-induced morphological changes, adhesion and motility, via its action on a specific substrate, paxillin and MAYP/PSTPIP2, a negative feedback regulator of macrophage and osteoclast production, in which we have described mutations that lead to an autoinflammatory disease in mice. Recently, utilizing gene expression, bioinformatic and cell biological approaches, we have shown that miR-21 is a novel CSF-1R pTyr-721–induced molecule that suppresses the macrophage M1 (inflammatory) phenotype and enhances their M2 (trophic) phenotype. In addition, we have shown that a novel ligand for the CSF-1R, interleukin-34 (IL-34), similarly activates the CSF-1R, but has a different tissue and cellular expression pattern from CSF-1.

Apart from our studies on CSF-1 and the CSF-1R in development and disease, we have also pioneered studies of the Shark tyrosine kinase in Drosophila, elucidating the pathways Shark regulates during the processes of embryonic dorsal closure (epithelial sheet movement) and the glial cell recognition and engulfment of dying cells.

In recent work in the nervous system, we have shown that all microglial development and maintenance is dependent on the CSF-1R and that the two CSF-1R ligands, CSF-1 and IL-34, also directly suppress self-renewal and enhance survival and differentiation of neural progenitor cells. We have identified protein tyrosine phosphatase zeta as an additional receptor for IL-34, but not CSF-1 and shown that the Csf1r^+/- mouse is a model of CSF-1 receptor-related leukoencephalopathy (CRL), formerly known as adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP), a dementia caused by dominant, inactivating mutations in the kinase domain of the CSF1R gene and most often misdiagnosed as Alzheimer’s disease. Over the last 5 years, we have shown that Csf1r heterozygosity in microglia is sufficient for the development of mouse CRL, that the genes encoding CSF-2 and CSF-3 are overexpressed in the brains of CRL patients and that CSF-2 and CSF-3 directly promote microglial dysfunction and pathology of CRL mice. Our current studies are focused on elucidating the mechanisms underlying CRL development in the Csf1r^+/- mouse and developing novel therapeutic approaches.

Selected recent original work

Wang, Y., Yeung, Y.G., Langdon, W.Y. and Stanley, E.R. (1996) c-Cbl Is Transiently Tyrosine phosphorylated, Ubiquitinated, and Membrane-targeted following CSF-1 Stimulation of Macrophages. J. Biol. Chem. 271:17-20. PMID: 8550554

Yeung, Y.G., Soldera, S. and Stanley, E.R. (1998) A novel macrophage actin-associated protein (MAYP) is tyrosine phosphorylated following CSF-1 stimulation. J. Biol. Chem. 273:30638 30642. PMID: 9804836

Lee, P.S.W., Wang, Y., Dominguez, M.G., Yeung, Y-G., Murphy, M.A. Bowtell, D.D.L. and Stanley, E.R. (1999) The Cbl protooncoprotein stimulates CSF-1 receptor multiubiquitination and endocytosis, and attenuates macrophage proliferation. EMBO J. 18:3616-3628. PMID: PMC1171440

Fernandez, R., Takahashi, F., Liu, Z., Steward, R., Stein, D. and Stanley, E.R. (2000) The Drosophila Shark tyrosine kinase is required for embryonic dorsal closure. Genes and Development 14:604-614. PMID: PMC316420

Pixley, F.J., Lee, P.S.W., Condeelis, J. and Stanley, E.R. (2001) Protein tyrosine phosphatase phi regulates paxillin tyrosine phosphorylation and mediates Colony Stimulating Factor 1 induced morphological changes in macrophages. Mol. Cell. Biol. 21:1795-1809. PMID: PMC86738

Dai, X-M., Ryan, G.R., Hapel, A.J., Dominguez, M.G., Kapp, S., Sylvestre, V. and Stanley, E.R. (2002) Targeted disruption of the mouse CSF-1 receptor gene results in osteopetrosis, mononuclear phagocyte deficiency, increased splenic progenitor cell frequencies and reproductive defects. Blood 99:111-120. PMID: 11756160

Yeung, Y-G. and Stanley, E.R. (2003) Proteomic approaches to analysis of the early events in CSF-1 signal transduction. Molecular and Cellular Proteomics 2: 1143-1155. PMID: 12966146

Wyckoff, J., Wang, W., Lin, E.L., Wang, Y., Pixley, F.J., Stanley, E.R., Graf, T., Pollard, J.W., Segall, J. and Condeelis, J. (2004). A paracrine loop between tumor cells and macrophages is required for tumor cell migration in mammary tumors.  Cancer Res. 64(19):7022-7029. PMID: 15466195

Dai X-M, Zong XH, Sylvestre V and Stanley ER. (2004) Incomplete restoration of colony stimulating factor-1 (CSF-1) function in CSF-1-deficient Csf1op/Csf1op mice by transgenic expression of cell surface CSF-1. Blood 103: 1114-1123. PMID: 14525772

Dai, X-M., Zong, X-H., Akhter, M.P. and Stanley, E.R. (2004). Osteoclast deficiency results in disorganized matrix, reduced mineralization, and abnormal osteoblast behavior in developing bone.  J. Bone Min. Res. 19(9):1441-1451. PMID: 15312244

Chitu, V., Pixley, F.J., Yeung, Y.G., Macaluso, F., Condeelis, J and Stanley, E.R. (2005). The PCH family member MAYP/PSTPIP2 directly regulates F-actin bundling and enhances filopodia formation and motility in macrophages. Mol. Biol. Cell. 16(6):2947-2959. PMID: PMC1142438

Grosse, J., Chitu, V., Marquardt, A., Hanke, P., Schmittwolf, C., Zeitlmann, L., Schropp, P., Barth, B., Yu, P., Paffenholz, R., Stumm, G., Nehls M., and Stanley E. R. (2006). Mutation of mouse MAYP/PSTPIP2 causes a macrophage autoinflammatory disease. Blood 107(8):3350-3358. PMID: PMC1895761

Biswas, R., Stein, D. and Stanley, E.R (2006). Drosophila Dok is required for embryonic dorsal closure. Development 133(2):217-227. PMID: 16339186

Nandi, S., Akhter, M.P., Seifert, M.F., Dai, X-M. and Stanley, E.R. (2006). Developmental and functional significance of the CSF-1 proteoglycan chondroitin sulfate chain. Blood 107(2):786-795. PMID: PMC1895624

Zeigenfuss, J.S.*, Biswas, R.*, Avery, M.A., Sheehan, A.E., Hong, K., Yeung Y-G., Stanley, E.R.** and Freeman, M.R.** (2008). Draper-dependent glial phagocytic activity is mediated by Src and Syk family kinase signaling. Nature 453(7197):935-939. PMID: PMC2493287

Huynh, D., Dai, X-M., Nandi, S., Lightowler, S., Trivett, M., Chan, C-K., Bertoncello, I., Ramsay, R.G.  and Stanley, E.R. (2009). CSF-1 dependence of Paneth cell development in the mouse small intestine. Gastroenterology 137(1):136-144. PMID: PMC2706482

Patsialou, A., Wyckoff, J., Wang, Y., Goswami, S., Stanley, E.R. and Condeelis J.S. (2009). Invasion of human breast cancer cells in vivo requires both paracrine and autocrine loops involving the colony stimulating factor-1 receptor. Cancer Research 69(24):9498-9506. PMID: PMC2794986

Xiong, Y., Song, D., Cai, Y., Yu, W., Yeung, Y.G., and Stanley, E.R. (2010). A CSF-1 receptor phosphotyrosine 559 signaling pathway regulates receptor ubiquitination and tyrosine phosphorylation. J. Biol. Chem. 286(2):952-960. PMID: PMC3020780

Wei, S., Nandi, S., Chitu, V., Yeung, Y-G., Yu, W., Huang, M., Williams, L.T., Lin, H. and Stanley, E.R. (2010). Functional overlap but differential expression of CSF-1 and IL-34 in their CSF-1 receptor-mediated regulation of myeloid cells. J Leukoc. Biol. 88(3):495-505. PMID: 20504948

Ginhoux, F., Greter, M., Leboeuf, M., Nandi, S., See, P., Gokhan, S., Mehler, M.F., Ng, L.G., Stanley, E.R., Samokhvalov, I.M. and Merad, M. (2010) Fate mapping studies reveal that adult microglia derive from primitive macrophages. Science 330(6005):841-845. PMID: 20966214

Sampaio, N., Yu, W., Cox, D., Wyckoff, J., Condeelis, J., Stanley, E.R. and Pixley, F.J. (2011). Phosphorylation of Y721 of the CSF-1R mediates PI3K association to regulate macrophage motility and enhancement of tumor cell invasion. J. Cell Sci. 124(Pt 12):2021-2031. PMID: PMC3104034

Yu, W., Chen, J., X. Ying, Pixley, F.J., Yeung, Y.G., and Stanley, E.R. (2012). Macrophage proliferation is regulated through CSF-1 receptor tyrosines 544, 559 and 807. J. Biol.Chem. 287:13694-704. PMID: 22375015.

Nandi, S., Gokhan,S., Dai, X-M., Wei, S., Enikolopov, G., Lin,H., Mehler, M.F.  and Stanley E.R. (2012). The CSF-1 receptor ligands IL-34 and CSF-1 exhibit distinct developmental brain expression patterns and regulate neural progenitor cell maintenance and maturation. Developmental Biology 367:100-13. PMID: 22542597.

Chitu V., Nacu, V., Charles, J.F., Henne, W.M., McMahon, H.T., Nandi, S., Ketchum, H., Harris, R., Nakamura, M.C., Stanley, E.R. (2012) PSTPIP2 deficiency in mice causes osteopenia and increased differentiation of multipotent myeloid precursors into osteoclasts. Blood 120:3126-35. PMID: 22923495.

Nandi, S., Cioce, M., Yeung, Y.G., Nieves, E., Tesfa, L., Lin, H., Hsu, A.W., Halenbeck R., Cheng, H.Y., Gokhan, S., Mehler, M.F., Stanley, E.R.(2013).Receptor-type protein tyrosine phosphatase zeta is a functional receptor for interleukin-34. J. Biol. Chem. 288:21972-86. PMID: 23744080

Chitu, V., Gokhan, S., Gulinello, M., Branch C.A., Patil, M., Basu, R., Stoddart C., Mehler M.F.  and Stanley E.R.(2014) Phenotypic characterization of a Csf1r haploinsuficient mouse model of adult-onset leukodystrophy with axonal spheroids and pigmented glia (ALSP).  Neurobiology of Disease, 74C:219-228. PMID: 25497733

Caescu, C.I., Guo,X., Tesfa, L., Bhagat, T.D., Verma, A., Zheng, D. and Stanley, E.R. (2015). Colony stimulating factor-1 receptor signaling networks inhibit mouse macrophage inflammatory responses by induction of microRNA-21. Blood, . Blood, 125:e1-13. PMID:25573988

Chitu, V., Biundo, F., Shlager, G.G.L., Park, E.S., Wang, P., Gulinello, M.E., Gokhan, S., Ketchum, H.C., Saha, K., DeTure, M. A., Dickson, D.W., Wszolek, Z.K., Zheng, D., Croxford, A.L., Becher, B., Sun, D., Mehler M.F. and Stanley E.R. (2020) Microglial homeostasis requires balanced CSF-1/CSF-2 receptor signaling. Cell Reports, 30:3004-3019. PMID: 32130903

Biundo, F., Chitu, V., Shlager, G.G.L., Park, E.S., Gulinello, M.E., Saha, K., Ketchum, H.C., Fernandes, C., Gökhan, Ş., Mehler, M.F.  and Stanley E.R. (2021) Microglial reduction of colony stimulating factor-1 receptor expression is sufficient to confer adult onset leukodystrophy. Glia, 69:779-791. PubMed PMID: 33079443

Selected recent reviews

Pixley, F.J. and Stanley, E.R. (2004) CSF-1 Regulation of the Wandering Macrophage: Complexity in action. Trends in Cell Biol. 14:628-638. PMID: 15519852

Chitu, V. and Stanley E. R. (2006).  Colony stimulating factor-1 in immunity and inflammation.  Current Opinion in Immunology 18(1):39-48. PMID: 16337366

Chitu, V. and Stanley, E.R. (2007). Pombe Cdc15 homology (PCH) proteins: coordinators of membrane-cytoskeletal interactions. Trends in Cell Biol. 17(3):145-156. PMID: 17296299

Stanley, E.R. (2009), “Colony Stimulating Factor-1 Regulation of Macrophages in Development and Disease” in Gordon, S. ed.) Innate Immunity: Host recognition and response in health and disease, The Biomedical & Life Sciences Collection, Henry Stewart Talks Ltd. London (online at http://www.hstalks.com/bio).

Stanley, E.R. and Chitu, V. (2014). CSF-1 Receptor Signaling in Myeloid Cells. Cold Spring Harb Perspect Biol. 6(6):1-21 PMID: 24890514.

Chitu, V. and Stanley, E.R. (2015). CSF-1 Receptor. In: The PDGF Receptor Family. Chapter 10 of Receptor Tyrosine Kinases: Family and Subfamilies. (D.L. Wheeler and Yarden, Y., eds.), Humana Press, New York. pp 373-538.

Caescu, C.I. and Stanley, E.R. (2015). FLT3. In: The PDGF Receptor Family. Chapter 10 of Receptor Tyrosine Kinases: Family and Subfamilies. (D.L. Wheeler and Yarden, Y., eds.), Humana Press, New York. pp 373-538.

Chitu,V., Gokhan,S., Nandi, S., Mehler, M. and Stanley, E.R. (2016) Emerging roles for the CSF-1 receptor and its ligands in the nervous system. Trends in Neuroscience, 39:378-393.

Chitu,V. and Stanley, E.R. (2017) Regulation of embryonic and postnatal development by the CSF-1 receptor. Curr. Top. Dev. Biol. 123:229-275.

Chitu, V., Gokhan, S., Nandi, S., Mehler, M. and Stanley, E.R. (2016) Emerging roles for the CSF-1 receptor and its ligands in the nervous system. Trends in Neuroscience, 39:378-393.

Chitu, V., Gökhan, Ş. and Stanley, E.R. (2021) Modeling CSF-1 receptor deficiency diseases – how close are we? FEBS J., Jun 19. doi: 10.1111/febs.16085. Online ahead of print. PMID: 34145972

Chitu, V., Biundo, F. and Stanley, E.R. (2021) Colony Stimulating Factors in the Nervous System. Semin Immunol. 54:101511. doi: 10.1016/j.smim.2021.101511. Epub 2021 Nov 4. PMID: 34743926

(last updated 08/02/22)