APPLICATION OF TECHNOLOGY:

• Drug discovery assays for developing therapies for allergic and atopic diseases and conditions, e.g. asthma, Crohn’s disease, myeloproliferative disorder, 5q31-associated leukemias and multiple sclerosis, and other cytokines-associated inflammatory disorders
  
• Association studies and single nucleotide polymorphisms (SNP) discovery
  
• DNA testing for regulators of allergic and atopic genes
  
• Discovering potential therapeutic targets
  

ADVANTAGES:

• Enhances the ability to diagnose allergic and atopic diseases
  
• Maps out research path for developing therapies that target the genetic roots of immune disorders instead of just symptoms
  

ABSTRACT: Edward Rubin and Kelly Frazer have identified and functionally characterized the conserved non-coding sequence (CNS)-1 as a potent regulator controlling the expression of three genes associated with numerous allergic and atopic diseases. Since regulatory sequence variation can contribute to human disorders as much as coding variation, this invention has important therapeutic and diagnostic applications.

The newly discovered sequences can be used to diagnose altered CNS sequences that increase or decrease the expression of IL-4, IL-13, and IL-5. Drs. Rubin and Frazer have also designed drug discovery assays and transgenic animals for developing therapies for allergic diseases and conditions. In addition, these sequences can be used to discover potential therapeutic targets by helping identify trans-acting factors that regulate cytokine expression via CNS-1.

Previous studies have identified several cytokine genes clustered at human 5q31 as biomedically important. These genes, interleukin-4 (IL-4), interleukin-13 (IL-13), and interleukin-5 (IL-5), while rapidly evolving, have remained physically close to each other in all species studied. IL-4, IL-13, and IL-5 are also coordinately co-activited in T helper 2-type (T_H2) cells. These observations suggest that these cytokines are clustered due to commonly shared, and previously unidentified, cis-regulatory sequences.

Using comparative analysis between humans, several mammals, chicken, and fish, Berkeley researchers identified the regulatory sequences associated with IL-4, IL-13, and IL-5 expression. In addition, the biological properties of CNS-1 were characterized through the creation and analysis of multiple lines of mice bearing a 450-kilobase human yeast artificial chromosome (YAC) transgene either containing or lacking the CNS-1 element. The results suggest that CNS-1 increases the likelihood that the human 5q31 T_H2 cytokines will be expressed, but does not act as a silencer or enhancer and therefore may be involved in modulating chromatin structure.

Expression of human cytokines in paired CNS-1wt and CNS-1del transgenics and control FVB mice. Naïve CD4+ T cells were stimulated with TH 2 conditions and analyzed on days 2, 3, 4, and 7 for expression of (A) human IL-4, (B) human IL-13, and (C) human IL-5. In the absence of CNS-1 the expression of these human cytokines was dramatically reduced.

STATUS: U.S. Patent #6,891,031; available for licensing

REFERENCE NUMBER: IB-1584

PUBLICATION:Loots, G.G., Locksley, R.M., Blankespoor, C.M., Wang, Z.E., Miller, W., Rubin, E.M., Frazer, K.A., “Identification of a Coordinate Regulator of Interleukins 4, 13, and 5 by Cross-Species Sequence Comparisons,” Science 2000, 288, 136-140.

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