Project Details
Description
DESCRIPTION (Adapted from applicant's abstract): Neurohematopoietic
modulation is beginning to be unravelled. This understanding has lead
to the addition of neuropeptides, such as the mammalian tachykinins to
the list of hematopoietic regulators. The mechanisms by which
neuropeptides regulate hematopoiesis will add to the general
understanding of the pathophysiology of diseases such as anemias and
leukemias. Substance P (SP) and neurokinin A (NK-A), members of the
tachykinins, modulate hematopoiesis partly through cytokine induction
in bone marrow (BM) stroma. Studies with specific antagonists indicate
that SP/NK-A interact with two (NK-1 and NK-2) of the three cloned
neurokinin receptors (NK-R) in stromal cells. IL-1 and c-kit ligand
modulate the numbers and affinities of SP receptors (SP-R) in stroma.
Amplification with specific NK-1 primers indicated that the size of
fragments induced by these cytokines are different than predicted. The
change in binding affinities as well as difference in fragment size
suggest that at least for NK-1, BM stroma may have different subtypes.
Transcriptional regulation of the NK-Rs will help to elucidate the
mechanisms by which the tachykinins regulate hematopoiesis. However,
this will only be possible if the NK-R cDNA in BM stroma is cloned.
This application will focus on cloning the NK-1 cDNA in human BM stroma
and elucidate its/their transcriptional regulation by specific cytokines
and neurotrophic factors. The working hypothesis is based on the
abilities of cytokines and neurotrophic factors found within the BM to
induce NK-1 in BM stroma. Since unstimulated stroma failed to amplify
visible NK-1 specific DNA fragments, in aim 1, the investigators will use
a cDNA library prepared from IL-1 stimulated BM stroma to clone the NK-1
cDNA. Specific probes, designed from the cloned cDNA(s) will be used
to identify the inducers (cytokines/neurotrophic factors) of NK-1
subtype(s). The regions within the NK-1 gene that are sensitive to these
'inducers' will be identified in aim 2. To address this question, the
sequence of the NK-1 promoter is required and this will be obtained after
cloning of the human NK-1 gene from a commercially available genomic
library. Different upstream sequences of the NK-1 gene will be used in
aim 3 to prepare heterologous reporter constructs which will be
transfected in immune and non- immune cell lines, BM stroma, and specific
stromal cells. Transfected cells will be stimulated with
cytokines/neurotrophic factors and reporter activities used to determine
activation. The answers obtained in this application for a FIRST award
will lead to the long term goal of this project which will include
studies of intracellular signalling pathway by NK-Rs in specific BM cell
populations and identification of transcriptional factors pertinent to
the NK-R gene.
Status | Finished |
---|---|
Effective start/end date | 8/1/97 → 7/31/02 |
Funding
- National Heart, Lung, and Blood Institute: $129,420.00
ASJC
- Genetics
- Oncology
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