RESUMO
Noninvasive and nonradioactive imaging modality to track and image apoptosis during chemotherapy of triple negative breast cancer is much needed for an effective treatment plan. Phosphatidylserine (PS) is a biomarker transiently exposed on the outer surface of the cells during apoptosis. Its externalization occurs within a few hours of an apoptotic stimulus by a chemotherapy drug and leads to presentation of millions of phospholipid molecules per apoptotic cell on the cell surface. This makes PS an abundant and accessible target for apoptosis imaging. In the current work, we show that PS monoclonal antibody tagged with indocyanine green (ICG) can help to track and image apoptosis using multispectral optoacoustic tomography Assuntos
Anticorpos Monoclonais/farmacocinética
, Apoptose/efeitos dos fármacos
, Verde de Indocianina/farmacocinética
, Imagem Óptica/métodos
, Fosfatidilserinas/farmacocinética
, Técnicas Fotoacústicas/métodos
, Neoplasias de Mama Triplo Negativas
, Animais
, Anticorpos Monoclonais/farmacologia
, Antineoplásicos/farmacocinética
, Antineoplásicos/farmacologia
, Feminino
, Verde de Indocianina/farmacologia
, Camundongos
, Camundongos Nus
, Fosfatidilserinas/farmacologia
, Neoplasias de Mama Triplo Negativas/diagnóstico por imagem
, Neoplasias de Mama Triplo Negativas/metabolismo
, Neoplasias de Mama Triplo Negativas/patologia
RESUMO
The slit diaphragm and the apical and basal membrane domains of podocytes are connected to each other by an actin-based cytoskeleton critical to the maintenance of the glomerular filtration barrier. In an effort to discover novel regulatory proteins of the podocyte foot process, we identified and characterized pdlim2, a member of the actin-associated LIM protein subfamily of cytosolic proteins typified by an N-terminal PDZ domain and a C-terminal LIM domain. In the kidney, the pdlim2 protein is highly specific for the glomerulus and podocyte foot processes as shown by RT-PCR, western blotting, immunofluorescence, and immunoelectron microscopy. In cultured podocytes, pdlim2 was associated with stress fibers and cortical actin. Pdlim2 seems to regulate actin dynamics in podocytes since stress fibers were stabilized in its presence. Mechanistically, pdlim2 interacts with two actin-associated podocyte proteins, α-actinin-4 and angiomotin-like-1, as shown by immunoprecipitation and yeast two-hybrid analyses. By semi-quantitative immunoelectron microscopy, there was a reduced expression of pdlim2 in podocytes of patients with minimal change nephrotic syndrome and membranous nephropathy, whereas its expression was unchanged in patients with focal segmental glomerulosclerosis. Hence, pdlim2 is a novel actin-regulating protein of podocyte foot processes that may have a role in the pathogenesis of glomerular diseases.