Project Ubermensch

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Project Ubermensch
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A Leap Towards Regeneration
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Regeneration

• Imagine a world
• Organs
• Limbs
• Spinal cord

Seems like science fiction But in Reality
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Many organisms can!

• Salamanders, zebrafish, planarian flatworms

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Salamander Limb Regeneration
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So, if they can do it why cant we?
We can but only up to a certain point
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The Basis of Regeneration

• Wound Epithelium
• Cellular dedifferentiation
• Naturally turn back cellular clock
• Proliferation

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Proposal

• Investigate genetic pathways and specific
  regulators of the dedifferentiation pathway in
  the salamander model
• Manipulation of specific regulators in
  orthologous human pathways ? dedifferentiation in
  human cells?

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The Importance

• Fundamental understanding of pathways give
  insight into control of dedifferentiated cells
• Regenerative pathways scar tissue vs.
  redevelopment- one piece to the puzzle

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The Impact

• Treatment for many diseases
• Help in tissue regeneration research
• Development of disease specific materials
• Insight into natural regenerative processes
  ?human regeneration

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Competing Technologies

• Induced pluripotent stem cells (Yu et al. 2007)
• Epigenetic reprogramming
• Clinical research of pluripotent stem cells

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• Knowns

• Unknowns

• Induce pluripotency in human somatic cells
• Nerve-dependent regeneration/ neurotrophic factor
  signaling (Kumar et al. 2007)

• Salamander genome
• Control of induced pluripotent stem cells -
  cancerous
• Degree of pluripotency
• Self assembly of cell lines into tissues

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Chitinase-erase

• Food Allergies
• When your immune system reacts against a harmless
  compound found in what you eat.

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Chitinase? What and Why?

• Plant defense Chintin-ases breakdown fungi and
  some bacterial cell walls
• Expressed in low amounts0.01 in grains is
  substantial
• Increase in production-exposure to chitin,
  chitosan, ethylene
• Increase Latex-Fruit syndrome due to our current
  ripening process with ethylene gas

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Importance Latex-Fruit Syndrome

• Chitinase causes Cross-reactive allergen with
  Latex
• -hevein-like domain at the N termius
• 1-6 of the population suffers from Latex allergy
• Latex-Fruit Allergy 30-50

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Importance Treatment of Severe Allergies

• Due to B cell/antibody sensitivity to specific
  antigens in a particular food.
• Recognition of Epitope
• Histamine release
• Anaphylatic shock
• Possible Death
• EpiPens a large dosage of Epinephrine

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Competing TechnologyMedicinally

• IgE Neutralization Omalizumab
• -monoclonal antibody
• -mainly for allergenic asthmatics
• Immunotherapy
• -exposing bits of allergenic substances to
  the body, increasing the concentrations over time
• -either shots or small bits of food

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Proposal

• Temporary Knockout of Chitinase before
  harvesting
• Control of Knockout through an environmentally
  sensitive promotor
• Indicator that confirms Chintinase knockout
• Feedback mechanism even after harvest

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Knowns Current Plant Technologies

• Insertion of Recombinant DNA-plants
• -Ti Plasmid
• -Gene gun
• Temporary knockout of proteins
• -RNAi
• -Transcription Repressors
• Plants have stem cellscan develop into an entire
  organism from a few cells

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Unknowns

• Chitinase protein production pathway
• Control of Promotor
• Developing an entire plant
• Progeny that does not need infection
• Would it be safe to eat? Taste the same?
• ? Accepted by Public?

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Impact

• Allergy-free or hypoallergenic foods more food
  options
• Same technology- eliminate or enhance certain
  chemicals in plants and even other organisms

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Want Milk?

• The purpose of this project is to allow lactose
  intolerant individuals to consume dairy products
  without having to ingest lactase with every
  ingestion of dairy.
• (1)

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Want Milk?

• Lactose intolerance affects 70-75 of people
  worldwide, with its prevalence varying by
  ethnicity.
• (2)

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Want Milk?

• General Plan of Action
• -Create a gene that codes for lactase and insert
  it into Bifidobacterium Bifidum
• -The bacteria will then begin producing lactase
• -Find a way to insert new bacteria into GI tract,
  such as direct ingestion.
• Potential medium yogurt
• -Possibility of bacterial conjugation

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Want Milk?

• This project would have a great impact worldwide
  if it was successful.
• Only 10 of Americans suffer from lactose
  intolerance, but success would improve the lives
  of over 70 of the worlds population

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Want Milk?

• Competition
• Lactaid and other brands make lactase that can be
  ingested with a meal, as well as lactose-free
  dairy products.
• (3)

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Want Milk?

• Knowns
• -Lack of ability to produce lactase causes
  lactose intolerance
• -B. Bifidum is beneficial to our health and
  digestion, is found in almost everyone, stays in
  the lower GI tract, is immobile, and safe to use
• -Lactase can be/has been produced through
  biological methods

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Want Milk?

• Unknowns
• -Whether the B. Bifidum would stay in the GI
  tract once it is introduced
• -What signals to use to trigger and halt enzyme
  production
• -Whether it is possible for the bacteria to
  produce sufficient amounts of lactase

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Sources

• A Leap Towards Regeneration?
• Resources
• Picture http//artfiles.art.com/5/p/LRG/8/892/IE
  NJ000Z/tree-frog.jpg tree frog (Intro slide)
• Pictures newswiseblog.blogspot.com/2009_05_01_arc
  hive.html Background of Earth
• www.bountyfishing.com Picture of axolotl
• http//webscript.princeton.edu/icouzin/website/wp
  -content/uploads/image/zebra-fish-61290.jpg
  Picture of zebrafish
• http//www.mdc-berlin.de/en/news/2009/20090721-int
  ernational_research_team_seeks_to_unrav/index.html
  Picture of planarian flatworms
• http//animals.nationalgeographic.com/staticfiles/
  NGS/Shared/StaticFiles/animals/images/1024/axolotl
  .jpg Picture of axolotl
• http//th09.deviantart.net/fs24/300W/f/2008/010/f/
  1/__Blind_Salamander_Protocol___by_XOTV.jpg
  Picture of cartoon salamander
• http//www.foxnews.com/images/325524/1_61_stemcell
  s320.gif Picture of induced pluripotent stem
  cells
• Articles
• Nishikawa, S., R.A. Goldstein, and C.R. Nierras.
  "The Promise of Human Induced Pluripotent Stem
  Cells for Research and Therapy." Nature Reviews
  Molecular Cell Biology. 9.Sept. (2008) 725-729.
• Yu, J. et al. "Induced Pluripotent Stem Cell
  Lines Derived from Human Somatic Cells." Science.
  318.Dec. (2007) 1917-1920.
• Kragl, M. et al. "Cells Keep a Memory of Their
  Tissue Origin During Axolotl Limb
  Regeneration." Nature. 460.July (2009) 62-66.
• Kumar, A. et al. "Molecular Basis for the Nerve
  Dependence of Limb Regeneration in an Adult
  Vertebrate." Science. 318.Nov (2007) 772-777.

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Sources continued

• Chitinase-erase
• Picture of Payaya (intro slide)
  http//www.thedailygreen.com/cm/thedailygreen/imag
  es/xw/papaya-clean-FD-lg.jpg
• Picture of Xolair (Omalizumab)
  http//2.bp.blogspot.com/_otrvz2aDOLk/R1F3Uc_hYjI/
  AAAAAAAAAB8/0JqWKUDnQB0/s1600-R/Xolair.jpg
• Picture of Latex glove http//en.wikipedia.org/wi
  ki/FileDisposable_gloves_09.JPG
• Picture of avocado http//en.wikipedia.org/wiki/F
  ileAvacado444.jpg
• Picture of N terminus hevein domain
  http//dmd.nihs.go.jp/latex/cross-e.html
• Picture of plant callus http//upload.wikimedia.o
  rg/wikipedia/commons/thumb/1/18/Callus1.jpg/220px-
  Callus1.jpg
• Picture of GUS reporter http//upload.wikimedia.o
  rg/wikipedia/commons/thumb/9/9c/Rice_embryo.png/22
  0px-Rice_embryo.png
• http//books.google.com/books?hlenlridMxFn0gN
  qEWMCoifndpgPA157dq22Gooday2222Agressiv
  eanddefensiverolesforchitinases22otsyitZdT
  SGMHsig6C0KXm93mM3NLu2a0-R9JWIUx_0vonepageq
  ffalse
• Articles
• Paul P. Belliveau. "Omalizumab A Monoclonal
  Anti-IgE Antibody." MedGenMed. 2005 7(1) 27.
• Elizabeth Landau. 'Exciting' advance reported in
  peanut allergy therapy. CNN. February 19, 2010.
  http//www.cnn.com/2010/HEALTH/02/19/aaas.food.all
  ergies/index.html
• R. A. Jefferson. The GUS Reporter Gene System.
  Nature 342, 837 - 838 (14 December 1989).
• http//www.nature.com/nature/journal/v342/n6251/ab
  s/342837a0.html
• Wagner, S. and Breiteneder, H. The latex-fruit
  syndrome. Biochem. Soc. Trans., 6, 935-940
  (2002) http//www.ncbi.nlm.nih.gov/pubmed/12440950
  ?doptAbstract
• Website on Latex-Fruit Syndrome
  http//dmd.nihs.go.jp/latex/cross-e.html

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Sources continued

• Want Milk?
• Websites Visited
• http//en.citizendium.org/wiki/Bifidobacterium_bif
  idum
• http//www.textbookofbacteriology.net/normalflora.
  html
• http//www.wrongdiagnosis.com/l/lactose_intoleranc
  e/prevalence.htm
• Picture Credits
• (2)http//www.dsm.com/le/en_US/maxilact/html/lacto
  se_intolerance.htm
• (3)http//www.lactaid.com
• (1)http//oregonstate.edu/dept/foodsci/dairy/dairy
  _links.htm
• (Introduction) http//badanimalpuns.files.wordpre
  ss.com/2009/12/cow.jpg