Discovery of a Highly Conserved Peptide in the Iron Transporter Melanotransferrin that Traverses an Intact Blood Brain Barrier and Localizes in Neural Cells

The blood-brain barrier (BBB) hinders the distribution of therapeutics intended for treatment of diseases of the brain. Our previous studies demonstrated that that a soluble form of melanotransferrin (MTf; Uniprot P08582; also known as p97, MFI2, and CD228), a mammalian iron-transport protein, is an effective carrier for delivery of drug conjugates across the BBB into the brain and was the first BBB targeting delivery system to demonstrate therapeutic efficacy within the brain. Here, we performed a screen to identify peptides from MTf capable of traversing the BBB. We identified a highly conserved 12-amino acid peptide, termed MTfp, that retains the ability to cross the intact BBB undigested, distribute throughout the parenchyma, and enter endosomes and lysosomes within neurons, astrocytes and microglia in the brain. This peptide may provide a platform for the transport of therapeutics to the CNS, and thereby offers new avenues for potential treatments of neuropathologies that are currently refractory to existing therapies

Curcuphenol possesses an unusual histone deacetylase enhancing activity that counters immune escape in metastatic tumours

Curcuphenol, a common component of the culinary spices, naturally found in marine invertebrates and plants, has been identified as a novel candidate for reversing immune escape by restoring expression of the antigen presentation machinery (APM) in invasive cancers, thereby resurrecting the immune recognition of metastatic tumours. Two synthetic curcuphenol analogues, were prepared by informed design that demonstrated consistent induction of APM expression in metastatic prostate and lung carcinoma cells. Both analogues were subsequently found to possess a previously undescribed histone deacetylase (HDAC)-enhancing activity. Remarkably, the H3K27ac ChIPseq analysis of curcuphenol-treated cells reveals that the induced epigenomic marks closely resemble the changes in genome-wide pattern observed with interferon-γ, a cytokine instrumental for orchestrating innate and adaptive immunity. These observations link dietary components to modifying epigenetic programs that modulate gene expression guiding poised immunity

Identification of Trypanosome Proteins in Plasma from African Sleeping Sickness Patients Infected with T. b. rhodesiense

Control of human African sleeping sickness, caused by subspecies of the protozoan parasite Trypanosoma brucei, is based on preventing transmission by elimination of the tsetse vector and by active diagnostic screening and treatment of infected patients. To identify trypanosome proteins that have potential as biomarkers for detection and monitoring of African sleeping sickness, we have used a ‘deep-mining” proteomics approach to identify trypanosome proteins in human plasma. Abundant human plasma proteins were removed by immunodepletion. Depleted plasma samples were then digested to peptides with trypsin, fractionated by basic reversed phase and each fraction analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This sample processing and analysis method enabled identification of low levels of trypanosome proteins in pooled plasma from late stage sleeping sickness patients infected with Trypanosoma brucei rhodesiense. A total of 254 trypanosome proteins were confidently identified. Many of the parasite proteins identified were of unknown function, although metabolic enzymes, chaperones, proteases and ubiquitin-related/acting proteins were found. This approach to the identification of conserved, soluble trypanosome proteins in human plasma offers a possible route to improved disease diagnosis and monitoring, since these molecules are potential biomarkers for the development of a new generation of antigen-detection assays. The combined immuno-depletion/mass spectrometric approach can be applied to a variety of infectious diseases for unbiased biomarker identification

Chaperones and protein isomerases identified in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.

<p>Chaperones and protein isomerases identified in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.</p

Variant Surface Glycoproteins (VSGs) discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients confirmed to have late-stage human African trypanosomiasis.

<p>Variant Surface Glycoproteins (VSGs) discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients confirmed to have late-stage human African trypanosomiasis.</p

List of human plasma proteins with mass spectrometric molar intensities similar to those from the most abundant trypanosome proteins found in plasma from sleeping sickness patients.

*<p>Protein concentrations were retrieved from reference <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071463#pone.0071463-Hortin1" target="_blank">[19]</a>. Human proteins reported to be depleted by the LC20 IgY14 and Supermix LC10 columns <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071463#pone.0071463-Patel1" target="_blank">[26]</a> have been omitted from this table.</p

Summary of patient information.

<p>Plasma and CSF were collected from <i>T. b. rhodesiense</i>-infected patients confirmed to have late stage human African trypanosomiasis at Lwala Hospital in central Uganda.</p

ELISA detection of trypanosome antigens in plasma from <i>T. b. rhodesiense</i> -infected patients with late-stage human African trypanosomisis.

<p>All samples were tested in duplicate and the average response is shown. Error bars represent 1 standard deviation.</p