Replies to post #274082 on Avid Bioservices Inc (CDMO)

[biopharm]

Now Amyloid levels found different between TBI vs Alzheimer's

PS Targeting combo will be used to treat Alzheimers ... and it will be so much nicer to have biomarkers to scientifically prove it. Peregrine has the patents filed and can make updates to any biomarkers and which BP will have access to these early biomarkers to compare them to current, Alzheimers clinical trials they have going on. Look for Alzheimers trial that is stopped for analysis.... and as some become astronomically amazed that the protein pathways that would change via PS Targeting...the leverage is completely on Peregrines side.

Adam Boxer (UCSF) is one that will likely KNOW for a fact that PS Targeting will help Alzheimers so it will be interesting if he fails to bring forward info

From the old....

Formation of Amyloid Fibers Triggered by Phosphatidylserine-Containing Membranes†

Hongxia Zhao ,‡ Esa K. J. Tuominen ,‡ and Paavo K. J. Kinnunen *‡§
Helsinki Biophysics and Biomembrane Group, Institute of Biomedicine, P.O. Box 63 (Haartmaninkatu 8), University of Helsinki, FIN-00014 Helsinki, Finland, and Memphys-Center for Biomembrane Physics, Physics Department, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark
Biochemistry, 2004, 43 (32), pp 10302–10307
DOI: 10.1021/bi049002c
Publication Date (Web): July 20, 2004

Protein misfolding has been shown to be the direct cause of a number of highly devastating diseases such as Alzheimer's disease, Parkinson's disease, and Creutzfeldt-Jacob syndrome, affecting the aging population globally. The deposition in tissues of amyloid fibrils is a characteristic of all these diseases, and the mechanisms by which these protein aggregates form continue to be intensively investigated. In only a fraction of cases is an underlying mutation responsible, and accordingly, what initiates amyloid formation in vivo is the major question that is addressed. In this study, we show that membranes containing phosphatidylserine (PS), a negatively charged phospholipid, induce a rapid formation of fibers by a variety of proteins, viz., lysozyme, insulin, glyceraldehyde-3-phosphate dehydrogenase, myoglobin, transthyretin, cytochrome c, histone H1, and a-lactalbumin. Congo red staining of these fibers yields the characteristic light green birefringence of amyloid, and fluorescent lipid tracers further reveal them to include phospholipids. Our results suggest that PS as well as other acidic phospholipids could provide the physiological low-pH environment on cellular membranes, enhancing protein fibril formation in vivo. Interestingly, all the proteins mentioned above either are cytotoxic or induce apoptosis. PS-protein interaction could be involved in the mechanism of cytotoxicity of the aggregated protein fibrils, perturbing membrane functions. Importantly, our results suggest that this process induced by acidic phospholipids may provide an unprecedented and generic connection between three current major areas of research:? (i) mechanism(s) triggering amyloid formation, (ii) cytotoxicity of amyloidal protein aggregates, and (iii) mechanism(s) of action of cytotoxic proteins.

http://pubs.acs.org/doi/abs/10.1021/bi049002c

To the New... EXOSOMES will be everywhere, including Alzheimers

Ballroom D
Symposium session 1A - EV biogenesis I
Chairs: Clotilde Théry and Stephen J. Gould 13:15-14:00
Go to:
O-1A-1
ApoE regulates ESCRT-independent sorting on exosomes and endosomal amyloid formation

Guillaume Van Niel 1, Ptissam Bergam1, Aurelie Di Cicco2, Ilse Hurbain1, Alessandra Lo Cicero1, Cecile Fort1, Florent Dingli3, Marie-Claude Potier4, Leon Schurgers5, Damarys Loew3, Daniel Levy2 and Graça Raposo1
1Department of Cell Biology, Curie Institute, Paris, France; 2Department of Physical Chemistry, Curie Institute, Paris, France; 3Department of Mass Spectrometry, Curie Institute, Paris, France; 4CNRS UMR7225, INSERM U1127, UPMC, Institut du Cerveau et de la Moelle, Paris, France; 5Department of Biochemistry–Vascular Aspects, Faculty of Medicine, Health & Life Science, Maastricht University, Maastricht, The Netherlands

Introduction: Exosomes are generated within secretory multivesicular endosomes (MVEs) as intraluminal vesicles (ILVs). To serve specific cellular functions, notably once secreted, ILVs are enriched with defined sets of proteins by various and still elusive sorting mechanisms within MVEs. Pigment cells have tuned their MVEs to produce amyloid fibrils derived from the protein PMEL. For this purpose, PMEL – the first protein reported as an ESCRT-independent cargo – is sorted in a CD63-dependent manner on ILVs that likely serve as potential seeding platforms for PMEL amyloidogenesis. Contrary to amyloids such as those associated with Alzheimer's disease, PMEL amyloids are non-toxic and are functional as they serve as a scaffolding structure for the synthesis of melanin. To better understand the mechanisms exploited on ILVs to avoid potential toxicity during PMEL amyloidogenesis, we have used exosomes as reporters of these endosomal processes. Methods: For this purpose, we have characterized exosomes derived from pigment cells by cryo-electron microscopy, mass spectrometry and western blot. We then investigated the role of the intracellular counterparts of exosomes, ILVS in cell lines and in vivo using siRNA, western blotting and morphological analysis by electron microscopy. Results: Characterization of exosomes derived from pigment cells revealed the association of exosomes and ILVs with apolipoprotein E (ApoE) and lipoparticles. We could show that ApoE is targeted to endosomes in a CD63-dependent/ESCRT-independent manner and facilitates the ESCRT-independent sorting of PMEL amyloidogenic fragments onto ILVs. At the surface of ILVs, ApoE regulates the formation of mature fibrils in melanocytic cell lines and in pigment cells in vivo. Summary/conclusion: These results established a clear molecular mechanism for ESCRT-independent sorting of PMEL. Moreover, the novel evidence that lipoparticles are associated to exosomes provides a breakthrough that might be exploited to reconsider the respective roles of each extracellular particle in pathologies. Finally our study establishes a paradigm for the mechanism by which ApoE, the first genetic risk for early onset Alzheimer's disease, regulates the assembly of mature amyloid fibrils under benign and pathological conditions.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4408443/