Moreover, Silvestrol significantly depleted STAT3 protein levels without affecting STAT3 mRNA steady state levels (Supp. inhibition of eIF4A-responsive transcripts, such as STAT3, as a viable approach to alleviate cachexia. Introduction Cachexia, a multi-factorial disease characterized by acute muscle wasting and weight loss, accompanies various inflammatory diseases such as cancer, sepsis and AIDS1,2. The abnormal catabolic state found in cachectic patients arises from a combination of Trimethobenzamide hydrochloride complex metabolic changes and dysregulation of certain humoral factors3C5. Cachexia is the primary cause of ~22% of cancer-related deaths6 and has been known for decades as being a major influence on mortality rate in cancer patients. In spite of the relevance of this syndrome as a contributor to cancer-related deaths, there are no widely employed therapeutics that effectively alleviate this disease7. Despite the convoluted etiology of cachexia, some important mediators of its underlying pathophysiology have been identified. Accumulating c-Raf evidence depicts certain pro-inflammatory cytokines and their downstream effectors as playing pivotal roles in the onset of cancer cachexia7,8. For example, the concurrent signaling of interferon (IFN) and tumor necrosis factor (TNF) (IT) can synergistically elicit muscle wasting by stimulating the activity of transcription factors including STAT3 and the heterodimeric NF-B9C12. NF-B signaling in skeletal muscle upregulates the muscle-specific E3 ligase MURF-1 and induces a loss of proteins integral for muscle fiber formation and maintenance, such as MyoD and Myogenin7,9,11,13. Moreover, we have shown that NF-B can also mediate muscle wasting by collaborating with STAT3 to markedly increase the transcription of inducible nitric oxide synthase (iNOS), an enzyme that catalyzes the conversion of L-arginine to citrulline resulting in the release of nitric oxide (NO)7,9,10,12. Systemic interleukin-6 (IL-6) signaling is also crucial in inducing muscle wasting and has been shown to be involved in the pathophysiology of at least some models of cancer cachexia6,14C17. Chronic IL-6 exposure has been directly linked to the aberrant activation of autophagic and ubiquitin-proteasomal degradation systems in the muscle17. Furthermore, many studies have shown the importance of STAT3 in the muscle wasting process in a variety of IL-6-dependent models of cancer cachexia. These observations demonstrate that STAT3 is Trimethobenzamide hydrochloride essential in cachexia driven by a multitude of cytokines including IFN, TNF and IL-618C22. Attempts at interfering with cytokine signaling Trimethobenzamide hydrochloride to impede cachexia progression have included the use of antibodies targeting TNF or IL-6, however the success of these therapeutic approaches was very limited23,24. The disappointing outcomes in these trials could be due to the involvement of multiple distinct pathways, the cooperation of which is required for cachexia onset or due to redundancy in the downstream effectors of TNF and IL-6, such as STAT312. In light of these results, therapies that can Trimethobenzamide hydrochloride disrupt multiple pathways or target redundant factors downstream of these humoral factors may be a more fruitful approach to combatting cachexia. Disrupting the initiation of eukaryotic mRNA translation, including the rate-limiting recruitment of the 40S ribosome via the eIF4F complex, has been shown to have anti-immunogenic, anti-oncogenic and anti-cachectic effects25C27. Compounds such as silvestrol, pateamine A (PatA) and hippuristanol (Hipp) mediate these effects by inhibiting the function of eIF4A, a RNA helicase component of eIF4F that unwinds complex secondary structures in mRNAs28. These compounds are believed to act in this manner by perturbing the translation of specific set of mRNAs containing complex secondary structures in their 5 untranslated region (UTR) that hinder ribosomal recruitment27C31. Hipp is an allosteric inhibitor that prevents eIF4A binding to RNA32 whereas PatA and silvestrol deplete eIF4A from the eIF4F complex by causing eIF4A to clamp onto RNA33,34 thus disrupting the interplay between eIF4A and dependent transcripts35. In the past decade, these and Trimethobenzamide hydrochloride other compounds that target the eIF4F complex have received considerable attention, with several in preclinical development25. The anti-inflammatory effects of compounds that alter eIF4A function prompted us to investigate their impact in cancer cachexia. Previously, we reported that low doses of PatA prevents cytokine-induced muscle wasting both and in a C26-adenocarcinoma tumour induced mouse model of muscle wasting27. Without affecting general translation, we found that this low dose of PatA selectively disrupts the translation of iNOS mRNA by increasing its affinity to eIF4A,.