Studies on the role of BCR in Philadelphia chromosome-positive leukemias

It is well established that the chimeric Bcr-Abl oncoprotein resulting from fusing 3$\sp\prime$ ABL sequences on chromosome 9 to 5$\sp\prime$ BCR sequences on chromosome 22 is the primary cause of Philadelphia chromosome-positive (Ph$\sp1$) leukemias. Although it is clear that the cis-Bcr sequence present within Bcr-Abl is able to activate the tyrosine kinase activity and F-actin binding capacity of Bcr-Abl which is critical for the transforming ability of BCR-ABL, the biological role of normal BCR gene product (P160 BCR) remains largely unknown. The previous finding by our lab that P160 BCR forms stable complexes with Bcr-Abl oncoprotein in Ph$\sp1$-positive leukemic cells implicated P160 BCR in the pathogenesis of Ph$\sp1$-positive leukemias. Here, we demonstrated that P160 BCR physically interacts with P210 BCR-ABL and become tyrosine phosphorylated when co-expressed with P210 BCR-ABL in COS1 cells while no tyrosine phosphorylation of P160 BCR can be detected when it is expressed alone. The results suggest that P160 BCR is a target for the Bcr-Abl tyrosine kinase. Although we were unable to detect stable physical interaction between P160 BCR and P145 c-ABL (Ib) in COS1 cells overexpressing both proteins, P160 BCR was phosphorylated on tyrosine residues when co-expressed with activated tyrosine kinase of P145 c-ABL (Ib). In addition, studies of tyrosine phosphorylation of BCR deletion mutants and 2-dimensional tryptic mapping of in vitro phosphorylated wild type and mutant (tyrosine to phenylalanine) Bcr-Abl indicated that tyrosine 177, 283 and 360 of Bcr represent some of the phosphorylation sites. Even though the significance of tyrosine phosphorylation of residues 283 and 360 of Bcr has not been determined, tyrosine phosphorylation of residue 177 within Bcr-Abl has been reported to be critical for its interaction with Grb2 molecule and subsequent activation of Ras signaling pathway. Here, we further demonstrated that tyrosine 177 phosphorylated P160 BCR is also able to bind to Grb2 molecule suggesting the role of P160 BCR in the Ras signaling pathway.^ Surprisingly, using 3$\sp\prime$ BCR antisense oligonucleotide to reduce the expression of P160 BCR without interfering with the expression of BCR-ABL resulted in increased growth or survival of B15 cells and M3.16 cells expressing either P185 BCR-ABL or P210 BCR-ABL respectively. The results provided strong arguments that P160 BCR may function as a negative regulator for cell growth.^ Considering all these results, we hypothesize that P160 BCR negatively regulate cell growth and tyrosine phosphorylation of P160 BCR turns off its growth suppressor function and turns on its growth stimulatory function. We further speculate that Bcr-Abl oncoprotein in leukemia cells stably interacts with and constitutively phosphorylates portions of P160 BCR converting it into a growth stimulatory state. In normal cells, the growth suppressor effects of P160 BCR could only be transiently and conditionally switched to growth stimulatory action by a strictly regulated cellular tyrosine kinase such as c-ABL. The model will be further discussed in the text. ^

Something real: A social marketing approach to smoking prevention among Montevideo youth

Uruguay has some of the strictest tobacco-control laws in Latin America. Despite this, youth smoking rates in Uruguay are amongst the highest in South America. Thus, it is important to identify strategies to prevent youth smoking in Uruguay. The current qualitative research study sought to identify intrapersonal and socioenvironmental factors that are associated with smoking among middle school youth in Uruguay. It also sought to develop potential prevention strategies and media messages that would resonate with youth for a social media campaign. The study was grounded in social cognitive theory and the theory of reasoned action/planned behavior, among other behavioral science theories; anthropological perspectives were also considered. To achieve these goals, 29 group and individual structured interviews were conducted in two private middle schools catering to lower and higher SES youth in Montevideo, Uruguay during the summer of 2012. One hundred and three study participants, including students, parents, and teachers, were interviewed. The structured interviews were recorded, transcribed, translated, back translated, coded and analyzed. The study findings show that positive attitudes towards smoking (i.e. to be seen, to increase status, to ensure women's equality, to looking old, and to service as a rite of passage), delinquent behavior (i.e. transgression/deviant behavior), social norms that support smoking (i.e. peer pressure and modeling, group membership/sense of belonging, parental modeling, and family support), easy access and availability to tobacco (i.e. retails stores) were factors associated with youth smoking. Potential protective factors may include parental support, negative attitudes towards smoking, sports/music, and smoke-free environments. Because study participants are accustomed to government-sponsored strong countermarketing graphic imaging, study participants selected even stronger images and messages as the preferred way to receive tobacco prevention messages. Something Real ("Algo Real") was a theme that resonated with the participants and chosen as the name for the proposed campaign. This campaign was designed as a multiple component intervention that included mass, school base, and family based strategies to prevent tobacco use. Some intervention materials specific to these intervention components were developed to target relevant intrapersonal and socioenvironmental factors identified above. These materials will be tested in future pilot studies and larger scale evaluation with this population, outside the scope of this dissertation. ^