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Acetylcholinesterase-derived peptides and uses thereof

a technology of acetylcholinesterase and peptides, applied in the field of stem cell survival and expansion, can solve the problems of limited molecular pathway(s), incomplete understanding of the mechanisms responsible for initiating this adjustment, and limited capacity to stimulate proliferation

Inactive Publication Date: 2007-03-22
SOREQ HERMONA +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanisms responsible for initiating this adjustment are not fully understood.
Alone, their capacity to stimulate proliferation is limited.
However, the molecular pathway(s) leading from short to long term processes and which enable the adjustment to stressful stimuli, are not yet known.

Method used

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  • Acetylcholinesterase-derived peptides and uses thereof
  • Acetylcholinesterase-derived peptides and uses thereof
  • Acetylcholinesterase-derived peptides and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Hydrocortisone Elevates ACHE Gene Expression in Hematopoietic Stem Cells

[0497] This example relates to ACHE gene expression in hematopoietic stem cells and the influence of hydrocortisone thereon. The inventors have searched the extended promoter of the human ACHE gene (cosmid accession no. AF002993) for consensus motifs that may bind stress-associated and hematopoietic transcription factors. Two such clusters, one located 17 Kb upstream from the transcription start site, and another positioned at the first intron, were found to include motifs for AP1, NFkB, EGR-1 (as identified by a matrix search against the TransFac database, Heinemeyer et al., Nucleic Acids Res. 26, 364-370, 1998), interleukin-6 (IL6), with the consensus sequence CTGGG / AAA, glucocorticoid responsive element (GRE) half palindromic site, TGTTCT, and Stat-5, TTCCCAGAA or TT(C / A)(C / T)N(A / G) (G / T)AA (FIG. 1A). Of these, the latter two motifs are known to be actively involved in hematopoiesis (Darnell et al., Science ...

example 2

Expression of ARP in CD34+ Cells

[0505] The expression of ARP in CD34+ hematopoietic cells was evaluated by flow cytometry in whole cord blood and bone marrow from a patient with immune thrombocytopenic purpura (ITP), as demonstrated in FIG. 2A and FIG. 2B respectively. Bone marrow from ITP patients was chosen to study ARP expression in hematopoietic progenitor cells due to the high turnover of normal CD34+ in these patients. Cells were fixed and permeabilized with Fix and Perm (Caltag, Calif., US) and stained with monoclonal antibodies to CD34 conjugated to pycoerythrin (Beckton Dickinson, California, US) indicated as FL-2 and with highly specific rabbit anti-ARP antibodies followed by anti rabbit antibodies conjugated to fluorescein isothiocyanate, expression indicated as percentage of positive cells.

[0506] These findings demonstrate higher expression of AChE-R in proliferating hematopoietic progenitors from either newborns or individuals suffering from over-proliferation of bloo...

example 3

Readthrough AChE is Overproduced in the Myeloidogenic Mid-gestation Liver

[0507] To study the relevance of each of the AChEmRNA transcripts during development of the hematopoietic organs, in situ hybridization was performed on paraffin-embedded sections taken from human fetuses at different gestational ages. Consistent with the embryonic spatiotemporal shifts in blood cell forming tissues, we observed changes in the labeling intensity with the various probes used, in the aorta-gonad-mesonephric region (AGM), liver, spleen and bone marrow cells. FIG. 3A schematically presents the migration of hematopoiesis between the various blood cell forming tissues during fetal development. The top left of the figure represents a sagital section of a human embryo showing the hematopoietic organs—AGM (aorta-gonad-mesonephros), LIV (liver), SPL (spleen), and BM (bone marrow). The top right of the figure is a scheme of gestational shifts in hematopoietic processes which shows the relative intensity ...

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Abstract

The invention relates to a cell growth and / or differentiation regulatory peptide comprising a sequence of about 9 to about 150 amino acids derived from acetylcholinesterase amino acid sequence, preferably from the C-terminal region of acetylcholinesterase. The invention also relates to pharmaceutical compositions comprising the peptides, particularly for use in promoting survival of stem cells, promoting differentiation of stem cells, promoting growth of stem cells and / or promoting the growth-enhancing effect of a growth factor on stem cells, alone, or in combination with other growth factors. Of particular interest is the use of the peptides in the treatment of thrombocytopenia, post-irradiation conditions, post-chemotherapy conditions, or conditions following massive blood loss and promotion of neural progenitors in use for cell therapies aimed at restoring neural functions in diseased individuals. Further, the invention relates to antibodies against the peptides, inter alia for diagnostic use, for example, the diagnosis of stress-induced male infertility. The invention also relates to in vitro and in vivo methods for screening of drugs that affect the central nervous system, and are potential modulators of interactions between the “readthrough” form of acetylcholinesterase, AChE-R, the intracellular receptor RACK1 and the kinase PKC.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation-in-part (CIP) of U.S. patent application Ser. No. 09 / 998,042, filed Nov. 30, 2001, which is a continuation-in-part of PCT international application No. PCT / IL00 / 00311, filed May 31, 2000, which claims priority of Israeli Patent Application No. 130224, filed May 31, 1999 and Israeli Patent Application No. 131707, filed Sep. 2, 1999, incorporated herein by reference.STATEMENT AS TO FEDERALLY SPONSORED RESEARCH [0002] This work was supported by the US Army Medical Research and Material Command DAMD 17-99-9547 (July 1999-August 2004) and the Defense Advance Research Project Agency DARPA N66001-01-C-8015 (May 2001-May 2004). The US Government has certain rights in this invention.FIELD OF THE INVENTION [0003] The invention is directed to the field of stem cell survival and expansion. Specifically, the invention is directed at the stem cell survival and expansion effects of peptides derived from acetylcholine...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/567C07K16/40A61K38/00C12N9/18
CPCA61K38/00C12N9/18C07K2319/00C07K16/40
Inventor SOREQ, HERMONAELDOR, AMIRAMELDOR, SOFIADEUTSCH, VARDAGRISARU, DAN
Owner SOREQ HERMONA
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