A receptor that mediates osteoprotegerin ligand (OPGL)-induced osteoclast differentiation and activation has been identified via genomic analysis of a primary osteoclast precursor cell cDNA library and is identical to the tumor necrosis factor receptor (TNFR) family member RANK. The RANK mRNA was highly expressed by isolated bone marrow-derived osteoclast progenitors and by mature osteoclasts in vivo. Recombinant OPGL binds specifically to RANK expressed by transfected cell lines and purified osteoclast progenitors. Transgenic mice expressing a soluble RANK-Fc fusion protein have severe osteopetrosis because of a reduction in osteoclasts, similar to OPG transgenic mice. Recombinant RANK-Fc binds with high affinity to OPGL in vitro and blocks osteoclast differentiation and activation in vitro and in vivo. Furthermore, polyclonal Ab against the RANK extracellular domain promotes osteoclastogenesis in bone marrow cultures suggesting that RANK activation mediates the effects of OPGL on the osteoclast pathway. These data indicate that OPGL-induced osteoclastogenesis is directly mediated through RANK on osteoclast precursor cells.

We have cloned and characterized a human gene encoding TP2 (telomerase-associated protein 2), a protein with similarity to reverse transcriptases and the catalytic telomerase subunits from Saccharomyces cerevisiae and Euplotes aediculatus. Indirect immunofluorescence revealed that TP2 was localized to the nucleus. Using antibodies to endogenous and epitope-tagged TP2, we found that TP2 was associated specifically with human telomerase activity and the recently identified telomerase-associated protein TP1. Mutation of conserved residues within the reverse transcriptase domain of TP2 severely reduced associated telomerase activity. These results suggest that telomerase is an evolutionarily conserved multisubunit complex composed of both structural and catalytic subunits.

Investigators using anti-EpoR antibodies for immunoblotting and immunostaining have reported erythropoietin receptor (EpoR) expression in nonhematopoietic tissues including human tumors. However, these antibodies detected proteins of 66 to 78 kDa, significantly larger than the predicted molecular weight of EpoR (56-57 kDa). We investigated the specificity of these antibodies and showed that they all detected non-EpoR proteins. C-20 detected 3 proteins in tumor cell lines (35, 66, and 100 kDa). Sequences obtained from preparative gels had similarity to the C-20-immunizing peptide. The 66-kDa protein was a heat shock protein (HSP70) to which antibody binding was abrogated in peptide competition experiments. Antibody M-20 readily identified a 59-kDa EpoR protein. However, neither M-20 nor C-20 was suitable for detection of EpoR using immunohistochemical methods. We concluded that these antibodies have limited utility for detecting EpoR. Thus, reports of EpoR expression in tumor cells using these antibodies should be viewed with caution. (Blood.

Background Supportive oncology practice can be enhanced by integrating brief and validated electronic patient-reported outcome (ePRO) assessment into the electronic health record (EHR) and clinical workflow. Methods 636 women receiving gynecologic oncology outpatient care received instructions to complete clinical assessments through Epic MyChart, the EHR patient communication portal. PROMIS computer adaptive tests (CATs) were administered to assess fatigue, pain interference, physical function, depression, and anxiety. Checklists identified psychosocial concerns, informational and nutritional needs, and risk factors for inadequate nutrition. Assessment results, including PROMIS T-scores with documented severity thresholds, were immediately populated in the EHR. Clinicians were notified of clinically elevated symptoms through EHR messages. EHR integration was designed to provide automated triage to social work providers for psychosocial concerns, health educators for information, and dietitians for nutrition-related concerns. Results Of 4,042 MyChart messages sent, 3,203 (79%) were reviewed by patients. The assessment was started by 1,493 (37%) patients, and once started 93% completed (1,386 patients). Using first assessments only, 49.8% of patients who reviewed the MyChart message completed the assessment. Mean PROMIS CAT T-scores indicated a lower level of physical function and elevated anxiety compared to the general population. Fatigue, pain, and depression scores were comparable to the general population. Impaired physical functioning was the most common basis for clinical alerts, occurring in 4% of patients. Conclusions We used PROMIS CATs to measure common cancer symptoms in routine oncology outpatient care. Immediate EHR integration facilitated the use of symptom reporting as the basis for referral to psychosocial and supportive care.

The circadian clock links our daily cycles of sleep and activity to the external environment. Deregulation of the clock is implicated in a number of human disorders, including depression, seasonal affective disorder, and metabolic disorders. Casein kinase 1 epsilon (CK1) and casein kinase 1 delta (CK1␦) are closely related Ser-Thr protein kinases that serve as key clock regulators as demonstrated by mammalian mutations in each that dramatically alter the circadian period. Therefore, inhibitors of CK1␦/ may have utility in treating circadian disorders. Although we previously demonstrated that a pan-CK1␦/ inhibitor, 4-[3-cyclohexyl-5-(4-fluoro-phenyl)-3H-imidazol-4-yl]-pyrimidin-2-ylamine (PF-670462), causes a significant phase delay in animal models of circadian rhythm, it remains unclear whether one of the kinases has a predominant role in regulating the circadian clock. To test this, we have characterized 3-(3-, a novel and potent inhibitor of CK1 (IC 50 ϭ 32 nM) with greater than 20-fold selectivity over CK1␦. PF-4800567 completely blocks CK1-mediated PER3 nuclear localization and PER2 degradation. In cycling Rat1 fibroblasts and a mouse model of circadian rhythm, however, PF-4800567 has only a minimal effect on the circadian clock at concentrations substantially over its CK1 IC 50 . This is in contrast to the pan-CK1␦/ inhibitor PF-670462 that robustly alters the circadian clock under similar conditions. These data indicate that CK1 is not the predominant mediator of circadian timing relative to CK1␦. PF-4800567 should prove useful in probing unique roles between these two kinases in multiple signaling pathways.All living things, from fungi to humans, have regular cycles aligning them with the daily events in their environment. These cycles, known as circadian rhythms, are controlled in mammals by the master clock located in the suprachiasmatic nucleus of the hypothalamus (Antle and Silver, 2005;Gallego and Virshup, 2007). At the cellular level, the molecular events behind clock cycling are described by the regular increase and decrease in mRNAs and proteins that define feedback loops, resulting in approximately 24-h cycles. The suprachiasmatic nucleus is primarily regulated, or entrained, directly by light via the retinohypothalamic tract. The cycling outputs of the suprachiasmatic nucleus, not fully identified, regulate multiple downstream rhythms, such as those in sleep and awakening, body temperature, and hormone secretion (Schibler et al., 2003;Ko and Takahashi, 2006). As anyone who has experienced jet lag knows, misalignment of the internal clock with the external environment profoundly affects well being. Furthermore, diseases, such as depression, seasonal affective disorder, and metaArticle, publication date, and citation information can be found at

A novel cyclin gene was discovered by searching an expressed sequence tag database with a cyclin box profile. The human cyclin E2 gene encodes a 404-amino-acid protein that is most closely related to cyclin E. Cyclin E2 associates with Cdk2 in a functional kinase complex that is inhibited by both p27Kip1 and p21 Cip1 . The catalytic activity associated with cyclin E2 complexes is cell cycle regulated and peaks at the G 1 /S transition. Overexpression of cyclin E2 in mammalian cells accelerates G 1 , demonstrating that cyclin E2 may be rate limiting for G 1 progression. Unlike cyclin E1, which is expressed in most proliferating normal and tumor cells, cyclin E2 levels were low to undetectable in nontransformed cells and increased significantly in tumor-derived cells. The discovery of a novel second cyclin E family member suggests that multiple unique cyclin E-CDK complexes regulate cell cycle progression.The eukaryotic cell cycle is regulated by a family of cyclindependent kinases (CDKs) that are cyclically activated to trigger the different phases of the cell cycle in the correct order and at the right time. CDKs are regulated by a number of different proteins, including the cyclins that bind and activate the CDKs to form a serine/threonine kinase holoenzyme complex. In mammals, D-type cyclins in conjunction with cyclins E and A are required for cells to traverse G 1 and enter S phase (19,25,29). There are three members of the D-type cyclins, two members of the cyclin A family, and, to date, one mammalian cyclin E gene (33). The formation of distinct G 1 cyclin-CDK complexes regulates the temporal phosphorylation of specific substrates that drive cells through G 1 and into S phase. Cyclin-CDK complexes are negatively regulated by two families of CDK inhibitors (34). The Ink4 family of CDK inhibitors exclusively regulate D-type cyclin-CDK complexes, while the Kip/Cip family regulate D-type cyclins as well as CDK complexes comprised of cyclin E or A. p27Kip1 and p21 Cip1 bind and inhibit cyclin E-Cdk2 complexes by acting as competitive inhibitors for substrates and by preventing cyclin-activating kinase (CAK)-mediated phosphorylation and activation of Cdk2 (23,34).Cyclin E was originally discovered by its ability to complement the G 1 cyclins in budding yeast (16,18). This observation suggested that cyclin E regulated the progression of cells through the cell cycle. It was later shown that cyclin E-Cdk2 complexes stimulate mammalian cells to traverse G 1 and enter S phase by the temporal phosphorylation of specific substrates such as the retinoblastoma tumor suppressor protein (Rb) (17,20,33). In fission yeast, a critical size must be reached prior to entry into mitosis, and this is normally regulated by coupling Cdk1 activation to cell growth. Premature activation of Cdk1 in fission yeast causes cells to enter mitosis at a reduced size. A similar effect occurs when cyclin E is overexpressed three-to fourfold in mammalian cells; the cells have a shorter G 1 and enter mitosis at a reduced size (25, 29). These resul...

In this study of patients with advanced refractory solid tumors, AMG 706 was well tolerated and displayed favorable pharmacokinetics and evidence of antitumor activity. Additional studies of AMG 706 as monotherapy and in combination with various agents are ongoing.

Context Lung cancer patients experience multiple, simultaneous symptoms related to their disease and treatment that impair functioning and health-related quality of life (HRQL). Computer technology can reduce barriers to nonsystematic, infrequent symptom assessment and potentially contribute to improved patient care. Objectives To evaluate the efficacy of technology-based symptom monitoring and reporting in reducing symptom burden in patients with advanced lung cancer. Methods This was a prospective, multisite, randomized controlled trial (RCT). Two hundred fifty-three patients were enrolled at three sites and randomized to monitoring and reporting (MR) or monitoring alone (MA). Patients completed questionnaires at baseline, 3, 6, 9 and 12 weeks and symptom surveys via interactive voice response (IVR) weekly for 12 weeks. MR patients’ clinically significant symptom scores generated an e-mail alert to the site nurse for management. The primary endpoint was overall symptom burden; secondary endpoints included HRQL, treatment satisfaction, symptom management barriers, and self-efficacy. Results This RCT failed to demonstrate efficacy of symptom monitoring and reporting in reducing symptom burden compared with monitoring alone in lung cancer. HRQL declined over 12 weeks in both groups (P<0.006 to P<0.025); at week 12, treatment satisfaction was higher in MA than MR patients (P<0.012, P<0.027). Adherence to weekly calls was good (82%) and patient satisfaction was high. Conclusion Feasibility of using a technology-based system for systematic symptom monitoring in advanced lung cancer patients was demonstrated. Future research should focus on identifying patients most likely to benefit and other patient, provider and health system factors likely to contribute to the system’s success.