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David Stanley Milstone, MD, PhD
Senior Scientist, Brigham and Women's Hospital
Assistant Professor of Pathology, Harvard Medical School

Brigham and Women's Hospital
Department of Pathology
75 Francis Street
Boston, MA 02115

Research Location: Harvard New Research Building

Research Email: milstone@rics.bwh.harvard.edu

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Research Narrative:

My primary research interest is developmental vascular biology and inflammation. We employ various types of stem cells, both in vitro and in vivo in mice to study the roles of vascular cells and their genes in formation, regression and function of blood vessels in normal and pathologic processes.

We developed one of the first experimental applications of embryonic stems cells (ESC) differentiated in vitro to study how and when during development endothelium becomes responsive to inflammatory cytokines. We discovered a sequential process during which endothelial adhesion molecule genes, upregulated during inflammation in adults, first show constitutive expression followed by cytokine responsiveness in utero. These results have important implications both for normal fetal development and for intrauterine and peripartum infections and inflammation of both the fetus and mother.

I created and analyzed one of the first lines of E-selectin-deficient mice, discovering an essential role for this inducible endothelial adhesion molecule in leukocyte firm adhesion and slow rolling during inflammation (collaboration with Dr. Gimbrone). I also observed, unexpectedly, that E-selectin, and specifically its cytoplasmic domain, is required for the antiangiogenic action of endostatin, an antitumor agent that acts by inhibiting blood vessels. I established E-selectin-deficient mice, in three genetic backgrounds, that The Jackson Laboratory now distributes for general scientific use. I also created ES cells bearing a cytoplasmic domain-deleted E-selectin for in vivo studies.

I used ESC approaches to create loss-of-function chimeric mice demonstrating that PPARγ is essential for adipocyte differentiation in vitro and formation of white and brown adipose tissue in vivo (collaboration with Drs. Mortensen, Spiegelman and Freeman). These studies also revealed that PPARγ, in addition to enhancing macrophage lipid uptake, also upregulates CD36 thus stimulating reverse cholesterol efflux. Understanding these opposing effects on intracellular lipid accumulation, with corresponding opposite effects on foam cell formation, helped resolve a significant controversy about the overall effect of PPARγ in atherosclerosis.

I used conditional mutations to show that placental failure causes PPARγ-deficient embryonic lethality, and to rescue mosaic mutant mice for postnatal studies. Collaborations studying these mosaic animals revealed unexpected roles for PPARγ in cardiac hypertrophy and insulin resistance. Other collaborations involved creating and studying mice with loss of function mutations of the heterotrimeric G-protein components Gαi2, Gαi3 and Gαo. and several mutations of vascular cell adhesion molecule 1 (VCAM-1). The latter revealed a predominant role for VCAM-1, compared to ICAM-1, in leukocyte recruitment during atherosclerosis.

My independent work, involving extensive experiments with trophoblast stem cells (TSC) in vitro plus analysis of these cells in vivo, shows that PPARγ is a critical regulator of TSC differentiation to form all mature trophoblast lineages. This involves cell autonomous transcriptional effects on lineage-determinate target genes plus complex intercellular differentiation pathways.

Finally, as supervisor of a Morphology Core within the Center for Excellence in Vascular Biology, I have developed and refined protocols for immunolocalization of proteins (immunohistochemistry) and mRNA (radioactive and non-radioactive in situ hybridization) in vascular tissues and cells. This core is an integral part of the Center, supporting several independent investigator projects and a Program Project Grant based entirely within the Center.
 


Education:
Washington University, 1985, MD
Washington University, 1985, PhD

Publications (Pulled from Harvard Catalyst Profiles):

1. Herrmann MD, Clunie DA, Fedorov A, Doyle SW, Pieper S, Klepeis V, Le LP, Mutter GL, Milstone DS, Schultz TJ, Kikinis R, Kotecha GK, Hwang DH, Andriole KP, Iafrate AJ, Brink JA, Boland GW, Dreyer KJ, Michalski M, Golden JA, Louis DN, Lennerz JK. Implementing the DICOM Standard for Digital Pathology. J Pathol Inform. 2018; 9:37.

2. O'Donnell PE, Ye XZ, DeChellis MA, Davis VM, Duan SZ, Mortensen RM, Milstone DS. Lipodystrophy, Diabetes and Normal Serum Insulin in PPAR?-Deficient Neonatal Mice. PLoS One. 2016; 11(8):e0160636.

3. Milstone DS, Ilyama M, Chen M, O'Donnell P, Davis VM, Plutzky J, Brown JD, Haldar SM, Siu A, Lau AC, Zhu SN, Basheer MF, Collins T, Jongstra-Bilen J, Cybulsky MI. Differential role of an NF-?B transcriptional response element in endothelial versus intimal cell VCAM-1 expression. Circ Res. 2015 Jul 03; 117(2):166-77.

4. Tache V, Ciric A, Moretto-Zita M, Li Y, Peng J, Maltepe E, Milstone DS, Parast MM. Hypoxia and trophoblast differentiation: a key role for PPAR?. Stem Cells Dev. 2013 Nov 01; 22(21):2815-24.

5. Rose JA, Rabenold JJ, Parast MM, Milstone DS, Abrahams VM, Riley JK. Peptidoglycan induces necrosis and regulates cytokine production in murine trophoblast stem cells. Am J Reprod Immunol. 2011 Sep; 66(3):209-22.

6. Duan SZ, Usher MG, Foley EL, Milstone DS, Brosius FC, Mortensen RM. Sex dimorphic actions of rosiglitazone in generalised peroxisome proliferator-activated receptor-gamma (PPAR-gamma)-deficient mice. Diabetologia. 2010 Jul; 53(7):1493-505.

7. Parast MM, Yu H, Ciric A, Salata MW, Davis V, Milstone DS. PPARgamma regulates trophoblast proliferation and promotes labyrinthine trilineage differentiation. PLoS One. 2009 Nov 30; 4(11):e8055.

8. Ye CP, Duan SZ, Milstone DS, Mortensen RM. G(o) controls the hyperpolarization-activated current in embryonic stem cell-derived cardiocytes. Am J Physiol Heart Circ Physiol. 2008 Feb; 294(2):H979-85.

9. Duan SZ, Christe M, Milstone DS, Mortensen RM. Go but not Gi2 or Gi3 is required for muscarinic regulation of heart rate and heart rate variability in mice. Biochem Biophys Res Commun. 2007 May 25; 357(1):139-43.

10. Duan SZ, Ivashchenko CY, Whitesall SE, D'Alecy LG, Duquaine DC, Brosius FC, Gonzalez FJ, Vinson C, Pierre MA, Milstone DS, Mortensen RM. Hypotension, lipodystrophy, and insulin resistance in generalized PPARgamma-deficient mice rescued from embryonic lethality. J Clin Invest. 2007 Mar; 117(3):812-22.