A wide role for NOTCH signaling in acute leukemia

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A wide role for NOTCH signaling in acute leukemia

• Cecilia Varnagy-Moskovitz
• Chiaramonte, Raffaella, A. Basile, E. Tassi, E.
  Calzavarra, V. Cecchinato, V. Rossi, A. Biondi,
  P. Comi. 2005, A wide role for NOTCH1 signaling
  in acute leukemia. Cancer Letters 219 113-120

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Definitions.

• Leukemia group of bone marrow diseases
  involving an uncontrolled increase in WBC.
  (Medical Encyclopedia).
• Acute fast development of the disease.
• Lymphocytic is develop from lymphocytes.
• Myelocytic develop from myelocytes.

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Definitions

• Hematopoiesis process of development of blood
  cells.
• Second messenger system where an intracellular
  signal is generated in response to an
  intercellular stimulation.
• Ligand molecule that binds to another.

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Definitions

• NOTCH gene family that encodes single pass
  trans-membrane receptors able to transduce
  intercellular signals involved in cells-fate
  determination. (Kojika).
• JAGGED1 important ligand for NOTCH signaling
  pathways.
• Signal pathway transference of information
  through a channel, mediated by second messengers.

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Symptoms of Leukemia.

• Fevers or night sweats.
• Headache.
• Bleeding.
• Swollen lymph nodes.

• Frequent infections.
• Feeling weak or tired.
• Pain in bones or joints.
• Weight loss.

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Diagnosis of leukemia.

• Physical exam.
• Blood test.
• Biopsy.
• Cytogenetics.
• Spinal tap.
• Chest X-ray.

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Treatment for Leukemia.

• Chemotherapy.
• Biological therapy.
• Bone marrow transplantation.

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A wide role for NOTCH1 signaling in acute leukemia

• Chiaramonte, Raffaella
• et al.

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NOTCH signaling and acute leukemia

• Basic terms we need to know
• T-acute lymphoblastic leukemia (T-ALL)
• Acute myelogenic leukemia (AML)
• B-cell precursor ALL (BCP-ALL)
• Delta/Serrate/ Lag2 ligands (DSL)

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NOTCH signaling facts.

• NOTCH pathway has a role in the process of
  hematopoiesis.
• NOTCH1 is related to the development of
  thymocites.
• NOTCH1 needs to be active in earliest stages of
  T-cell commitment in thymus to be able to develop
  T-lymphocytes.
• If this doesnt happen B-lineage will be
  developed.

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NOTCH signaling facts.

• NOTCH1 has also important role in signaling
  alterations in pathogenesis of T-ALL.
• T-ALL patients present a translocation that leads
  to over expression of NOTCH1
• NOTCH pathway is activated in T-ALL cell lines.

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Materials and Methods

• Pediatric patients
• 32 children with T-ALL
• 25 with BCP-ALL
• 20 with AML
• Cell lines used were T-ALL and B-cell derived
  leukemia/lymphoma.

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Materials and Methods.

• Extraction of RNA and retro-transcripted.
• PCR test and amplifications.
• Amplified DNAs separated by electrophoresis.
• Results analyzed by ANOVA (analysis of variance)
  test

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Question!!

• What is the gene expression involved in NOTCH
  pathway activity related with three groups of
  patients T-ALL, BCP-ALL, and AML???

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Hypothesis.

• Ho gene expression values are equal across the
  different cell types.
• H1 at least two cell types gene expression
  values are different.
• For rejection of H0 Plt0.01

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Test and results.

• Analysis of T-ALL cell lines and DSL ligands.
  (control lymphoma cell line)
• In all T-ALL cell lines expressed at least one
  ligand.
• Comparison of expression values of NOTCH related
  genes (NOTCH1, HES1, pT? , and JAGGED1)

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RT-PCR analysis of DSL ligands expression in
T-ALL cell lines.

• Control cells Normalization gene

Chiaramonte et al.
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RT-PCR analysis of NOTCH pathway related genes

Target genes Ligands Normalization
gene
Chiaramonte et al.
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RT-PCR analysis of NOTCH pathway related genes.










Target genes Ligands Normalization
gene
Chiaramonte et al.
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Comparison test

• To know which particular population differs from
  others.
• To have a significant difference Plt0.05.

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Comparison test.
Chiaramonte et al.
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Comparison test.
Chiaramonte et al.
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Results

• After comparison of the three groups
• NOTCH1 levels in T-ALL and AML were equivalent,
  but higher than BCP-ALL (Plt0.01)
• For HES1 and pT?, T-ALL present highest levels of
  expression, compared to AML and BCP-ALL.
• JAGGED1 is more frequent in AML than T-ALL and
  BCP-ALL.

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NOTCH role.
Chiaramonte et al.
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Conclusions.

• Cell fate is regulated by NOTCH1.
• NOTCH1 interferes in cell differentiation.
• NOTCH1 helps avoid apoptosis of T-cell lineage
  differentiation.
• T-ALL present high levels of NOTCH1
• High level of pathway activity is related to
  NOTCH ligands.

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Conclusions

• JAGGED1 expression related to differentiation in
  hematopoietic progenitors.
• JAGGED1 has no relation with NOTCH pathway.
• NOTCH1 has an important role in T-ALL.

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References.
1.- American Cancer Society. What is Acute
Myeloid Leukemia? January 01, 2005. February
2005. www.cancer.org 2.- Chiaramonte, R. et al.
Differential regulation of NOTCH signal
transduction in leukemia and lymphoma cells in
culture. J. Cell. Biochem. 88. 2003569-577. 3.-
Janeway et al. Immunobiology. Garland Science
Publishing. 2005. 4.- Kojika, S. et al.NOTCH
Receptors in Hematopoiesis. Exp. Hematol. 29.
2001 1041-1052. 5.- Medical Dictionary Online.
Signal Pathway. 2005. February 2005. lt
http//www.online-medical-dictionary.org/omd.asp?q
signalpathwaygt 6.- Medical Encyclopedia from
Medline Plus. Leukemia. June 02, 2003. February
2005. http//www.nlm.nih.gov/medlineplus/ency/arti
cle/001299.htm 7.- Milner, L.A. et al. NOTCH as a
Mediator of cell fate determination in
hematopoiesis evidence and speculation. Blood
93. 1999 2431-2444. February 2005.
lthttp//www.bloodjournal.org/cgi/content/full/93/8
/2431gt 8.- National Cancer Institute. What you
need to know about Leukemia. March 03, 2003.
February 2005. ltwww.cancer.govgt 9.- Pear, W.S. et
al. Exclusive development of T cell neoplasm in
mice transplanted with bone marrow expressing
activated NOTCH alleles. Exp. Med. 183.
19962283-2291. lthttp//www.jem.org/cgi/reprint/1
83/5/2283gt 10.- Radtke, F. et al. Deficient T
cell fate specification in mice with an induced
inactivation of NOTCH1. Immunity 10.
1999547-558. 11.- Shelly, L. et al. NOTCH-1
inhibits apoptosis in murine erythroleukemia
cells and is necessary for differentiation
induced by hybrid polar compounds. J. Cell.
Biochem. 73. 1999164. 12.- Washburn, T. et al.
NOTCH activity influences the alpha/beta versus
gamma/delta lineage decision. Cell 88.
1997833-843.  
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Thank you

• Questions

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