Quality of life after immune suppressive therapy in aplastic anemia.

Acquired aplastic anemia (AA) is a rare form of immune-mediated bone marrow failure, which can result in life-threatening infections or bleeding if left untreated. Treatment consists of either immune suppressive therapy (IST) or allogeneic stem cell transplantation (alloHSCT). While considerable research has been published regarding survival, response rate and toxicity of both treatments, knowledge on the impact on quality of life (QoL) is scarce. We used the recently developed AA-specific QoL questionnaire (QLQ-AA/PNH-54) to evaluate QoL in a single center cohort of AA patients who were successfully treated with IST. The 54 questions represent 12 different QoL domains. Results were analyzed for all patients and grouped based on hematologic response (complete response (CR) or partial response (PR)). Thirty-six successfully treated adult patients (15 in CR, 21 in PR) completed the questionnaire (median age 54 years, range 21-71; median time since last IST 5 years, range 0-41). Fatigue was experienced by 83% of patients. Even though total QoL scores did not significantly differ between patients with PR and CR (105 vs 92, p-value 0,17) there appeared to be a trend towards higher scores in patients with PR, especially in domains concerning psychological wellbeing. This trend was most clear in the domains fear of progression (2,12 in PR patients vs 1,73 in CR patients; p-value 0,08) and role functioning (2,22 vs 1,88; p-value 0,07). In conclusion, patients with AA continue to experience psychological and physical effects despite successful IST.

Wnt-5a expression in the rat neuronal progenitor cell line ST14A.

Transplantation of cells derived from embryonic stem cells is currently under investigation as a promising strategy to restore functional deficits in neurodegenerative diseases, e.g. Parkinson's disease. To generate cells suitable for transplantation, a neuronal progenitor cell line (ST14A) was derived from embryonic day 14 rat striatum by stable retroviral transfection of the temperature-sensitive SV40 large T antigen and genetically modified by transfection with constructs of the neurotrophic factors ciliary neurotrophic factor (CNTF) [CNTF-ST14A] and glial cell line-derived neurotrophic factor (GDNF) [GDNF-ST14A], respectively. In order to investigate the capacity of these cells to regulate neuronal growth and physiological differentiation, e.g. remodeling of axons and synaptogenesis, we analyzed the expression of molecules which control the cell fate during embryonic development. For the first time, we found endogenous Wnt-5a, a regulator molecule that can induce dopaminergic phenotype, by RT-PCR, Western blot and flow cytometry in the neuronal progenitor cell line ST14A and its derivatives CNTF-ST14A and GDNF-ST14A. The protein was transiently upregulated at the differentiation-inducing non-permissive temperature of 39 degrees C and it was also secreted into the culture medium. Our findings are based on in vitro investigation of artificially immortalized cell lines. However, they raise the possibility that neuronal progenitor cells that might be used to treat neurodegenerative diseases express Wnt-5a, thus promoting their potential for dopaminergic differentiation.

Overexpression of glial cell line-derived neurotrophic factor induces genes regulating migration and differentiation of neuronal progenitor cells.

The glial cell line-derived neurotrophic factor (GDNF) is involved in the development and maintenance of neural tissues. Mutations in components of its signaling pathway lead to severe migration deficits of neuronal crest stem cells, tumor formation, or ablation of the urinary system. In animal models of Parkinson's disease, GDNF has been recognized to be neuroprotective and to improve motor function when delivered into the cerebral ventricles or into the substantia nigra. Here, we characterize the network of 43 genes induced by GDNF overproduction of neuronal progenitor cells (ST14A), which mainly regulate migration and differentiation of neuronal progenitor cells. GDNF down-regulates doublecortin, Paf-ah1b (Lis1), dynamin, and alpha-tubulin, which are involved in neocortical lamination and cytoskeletal reorganization. Axonal guidance depends on cell-surface molecules and extracellular matrix proteins. Laminin, Mpl3, Alcam, Bin1, Id1, Id2, Id3, neuregulin1, the ephrinB2-receptor, neuritin, focal adhesion kinase (FAK), Tc10, Pdpk1, clusterin, GTP-cyclooxygenase1, and follistatin are genes up-regulated by GDNF overexpression. Moreover, we found four key enzymes of the cholesterol-synthesis pathway to be down-regulated leading to decreased farnesyl-pyrophospate production. Many proteins are anchored by farnesyl-derivates at the cell membrane. The identification of these GDNF-regulated genes may open new opportunities for directly influencing differentiation and developmental processes of neurons.