The endothelium functions as a semi-permeable barrier between the blood plasma and interstitium thus regulating tissue fluid homeostasis. Impairment of the endothelial barrier is a key early event in the development of Adult Respiratory Distress Syndrome (ARDS). This condition is clinically manifested as a severe loss of gas exchange capacity and hypoxemia that is often fatal.
The focus of my research is on the cross-talk between microtubule cytoskeleton and VE-cadherin-mediated adhesions in lung microvascular endothelial cells. Dynamic interaction between cytoskeleton and adherens junctions is known to be important for maintenance of basal endothelial barrier permeability and underlies changes in endothelial permeability in response to different mediators. I am investigating the molecular and signaling mechanisms regulating microtubule cytoskeleton downstream of adherens junction and how changes in microtubule dynamics affect integrity of endothelial monolayer in response to extracellular stimuli. By establishing how MTs mediate increased lung vascular permeability we will be in a position to define novel therapeutic targets to treat ARDS.
Y.A. Komarova, S.M. Vogel, A.B. Malik “IP3R derivative peptide prevents inflammation-induced pulmonary vascular leakage and lethality in sepsis”. USA Provisional Patent, DD060
EB1 and EB3 promote cilia biogenesis by several centrosome-related mechanisms. Schrøder JM, Larsen J, Komarova Y, Akhmanova A, Thorsteinsson RI, Grigoriev I, Manguso R, Christensen ST, Pedersen SF, Geimer S, Pedersen LB. J Cell Sci. 2011 Aug 1;124(Pt 15):2539-51
Caveolin-1-eNOS signaling promotes p190RhoGAP-A nitration and endothelial permeability. Siddiqui MR, Komarova YA, Vogel SM, Gao X, Bonini MG, Rajasingh J, Zhao YY, Brovkovych V, Malik AB. J Cell Biol. 2011 May 30;193(5):841-50.
AKAP9 regulation of microtubule dynamics promotes Epac1-induced endothelial barrier properties. Sehrawat S, Ernandez T, Cullere X, Takahashi M, Ono Y, Komarova Y, Mayadas TN. Blood. 2011 Jan 13;117(2):708-18. Epub 2010 Oct 15.
Phosphorylation controls autoinhibition of cytoplasmic linker protein-170. Lee HS, Komarova YA, Nadezhdina ES, Anjum R, Peloquin JG, Schober JM, Danciu O, van Haren J, Galjart N, Gygi SP, Akhmanova A, Borisy GG. Mol Biol Cell. 2010 Aug 1;21(15):2661-73. Epub 2010 Jun 2.
Komarova Y., Malik AB. Regulation of endothelial permeability via paracellular and transcellular transport pathways. Annu. Rev. Physiol. 72: 9.1-9.31. DOI: 10.1146/annurev-physiol-021909-135833. 2010.
Mahmud G, Campbell C J., Bishop K J M, Komarova YA, Chaga O, Soh S, Huda S, Kandere-Grzybowska K, Grzybowski B A. Directing cell motions on micropatterned ratchets. Nat. Physics 5(8): 606-612. DOI: 10.1038/nphys1306. 2009
Schober JM, Cain JM, Komarova YA, Borisy GG. Migration and actin protrusion in melanoma cells are regulated by Eb1 protein. Cancer Lett. 284(1): 30-6. 2009.
Komarova Y., De Groot CO., Grigoriev I., Gouveia SM, Munteanu EL, Schober JM, Honnappa S, Buey RM, Hoogenraad CC, Dogterom M, Borisy GG, Steinmetz MO, Akhmanova A. Mammalian end binding proteins control persistent microtubule growth. J Cell Biol. 184(5): 691-706. 2009. PMC268640
Komarova Y., Malik A.B. FGF signaling preserves the integrity of endothelial adherens junctions. Dev Cell. 15(3): 335-336. 2008. PMC2662495
Komarova YA, Mehta D., Malik AB. Dual regulation of endothelial junctional permeability. Sci STKE. 2007(412): re8. Review. 2007.
Schober J, Komarova YA, Chaga O, Borisy G. Microtubule-targeting dependent reorganization of filopodia. J Cell Sci. 120:1235-1244. 2007.
Komarova YA, Lansbergen G, Galjart N, Grosveld F, Borisy G, Akhmanova A. EB1 and EB3 control CLIP dissociation from the ends of growing microtubules. Mol. Biol. Cell. 16 (11):5334-5345. 2005.
Kandere-Grzybowska K, Campbell C, Komarova Y, Grzybowski B, Borisy G. Molecular dynamics imaging in micropatterned living cells. Nat. Methods. 2 (10):739-741. 2005.
Peloquin J, Komarova YA, Borisy G. Conjugation of fluorophores to tubulin. Nat. Methods 2: 1-5. 2005.
Lansbergen G, Komarova YA, Modesti M, Wyman C, Hoogenraad CC, Goodson HV, Lemaitre RP, Drechsel DN, Van Munster E, Gadella TW Jr, Grosveld F, Galjart N, Borisy GG, Akhmanova A. Conformational changes in CLIP-170 regulate its binding to microtubules and dynactin localization. J Cell Biol. 166:1003-1014. 2004.
Komarova YA, Bortorello AM, Smith K, Leibiger IB, Efandiev R, Pedemonte CH, Borisy GG, Sznajder JI. Analysis of Na+, K+-ATPase motion and incorporation into the plasma membrane in response to G protein-coupled receptor signals in living cells. Mol. Biol. Cell 14: 1149-1157. 2003.
Komarova YA, Akhmanova AS, Kojima S, Galjart N, Borisy GG. Cytoplasmic linker proteins promote microtubule rescue in vivo. J Cell Biol. 159: 589-599. 2002.
Komarova YA, Vorobjev IA, Borisy GG. Life cycle of MTs: persistent growth in the cell interior, asymmetric transition frequencies and effects of the cell boundary. J Cell Sci. 115: 3527-3539. 2002.
Komarova Y., J. Peloquin, and G. Borisy. 2004. Microinjection of Fluorophore–labeled Proteins In Live Cell Imaging: A Laboratory Manual. Robert D. Goldman, David L. Spector, editors, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 67-87.
Cytoskeleton dynamics in cells
Movie 1.MT dynamics in CHO-K1 cells. In the steady state, MTs grow persistently from the centrosome toward the cell margin (MTs growing from the centrosome are in red). The growth is compensated by shortening from both plus (MTs shortened from the periphery back to the centrosome are in blue) and minus (MTs released from the centrosome and shortened from the minus ends are in green) ends. Time is shown in the upper right corner in minutes and seconds. Bar, 5 µm. Images were acquired on a spinning disk confocal microscope system. The system was equipped with a Nikon Eclipse TE 200 inverted microscope (Plan Fluor 100x 1.3 NA objective); spinning disk confocal scane-head (Yokogawa Electronics Corp., Tokyo, Japan); 3W argon laser and with 12-bit digital cooled CCD Cool Snap HQ CCD camera (Photometrics, Tucson, AZ).
Movie 2.Dynamics of noncentrosomal MTs in PtK-1 cell. In PtK-1 cells, minus ends of noncentrosomal MTs (colorized in blue and red) are stable and display no dynamics. Plus ends undergo alternating phases of growth and shortening. Plus and minus ends of MTs are indicated with “+” and “–,” respectively. Images were collected at 5-second intervals. Bar, 5 µm.
Movie 3.Formation of noncentrosomal MTs in PtK-1 cell. In PtK-1 cells, noncentrosomal MTs are formed by a “de novo” mechanism (blue) and via the "breakage" of preexisting MT (red). Plus and minus ends of MTs are indicated with “+” and “–.” Images were collected at 5-second intervals. Bar, 5 µm. Images were acquired using the same system as that for Movie 1.
Movie 4. Dynamics of actin in a locomoting fish keratocyte. In locomoting fish keratocytes, actin “speckles” (circles) remain stationary with respect to the substratum while the leading edge is advancing. Images were collected at 2-second intervals. Bar, 5 µm. Images were acquired using the same system as that for Movie 1.