Project Ubermensch A Leap Towards Regeneration Regeneration Imagine

Project Ubermensch

A Leap Towards Regeneration

Regeneration Imagine a world … Organs Limbs Spinal cord Seems like science fiction But in Reality…

Many organisms can! Salamanders, flatworms zebrafish, planarian

Salamander Limb Regeneration mputa A b m i L Wound healing Blastema Formation Reprogramming and Redevelopment

So, if they can do it… why can’t we? We can… but only up to a certain point

The Basis of Regeneration • • Wound Epithelium Cellular dedifferentiation • Naturally turn back cellular clock • Proliferation

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?

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

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

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

Knowns Induce pluripotency in human somatic cells Nerve-dependent regeneration/ neurotrophic factor signaling (Kumar et al. 2007) Unknowns Salamander genome Control of induced pluripotent stem cells cancerous Degree of pluripotency Self assembly of cell lines into tissues

Chitinase-erase Food Allergies: When your immune system reacts against a harmless compound found in what you eat.

Chitinase? What and Why? Plant defense: Chintin-ases breakdown fungi and some bacterial cell walls Expressed in low amounts— 0. 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

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%

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 Epi. Pens: a large dosage of Epinephrine

Competing Technology: Medicinally • Ig. E 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

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

Knowns: Current Plant Technologies Insertion of Recombinant DNA-plants -Ti Plasmid -Gene gun Temporary knockout of proteins: -RNAi -Transcription Repressors Plants have stem cells—can develop into an entire organism from a few cells

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?

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

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)

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

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

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 world’s population

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

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

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

Sources A Leap Towards Regeneration? Resources Picture: http: //artfiles. art. com/5/p/LRG/8/892/IENJ 000 Z/tree-frog. jpg tree frog (Intro slide) Pictures: newswiseblogspot. com/2009_05_01_archive. 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/20090721 international_research_team_seeks_to_unrav/index. html Picture of planarian flatworms http: //animals. nationalgeographic. com/staticfiles/NGS/Shared/Static. Files/animals/images/1024/axolotl. j pg Picture of axolotl http: //th 09. deviantart. net/fs 24/300 W/f/2008/010/f/1/__Blind_Salamander_Protocol___by_XOTV. jpg Picture of cartoon salamander http: //www. foxnews. com/images/325524/1_61_stemcells 320. 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.

Sources continued… Chitinase-erase Picture of Payaya (intro slide) : http: //www. thedailygreen. com/cm/thedailygreen/images/xw/papaya-clean-FD-lg. jpg Picture of Xolair (Omalizumab): http: //2. bp. blogspot. com/_otrvz 2 a. DOLk/R 1 F 3 Uc_h. Yj. I/AAAAAB 8/0 Jq. WKUDn. QB 0/s 1600 -R/Xolair. jpg Picture of Latex glove: http: //en. wikipedia. org/wiki/File: Disposable_gloves_09. JPG Picture of avocado: http: //en. wikipedia. org/wiki/File: Avacado 444. jpg Picture of N terminus hevein domain: http: //dmd. nihs. go. jp/latex/cross-e. html Picture of plant callus: http: //upload. wikimedia. org/wikipedia/commons/thumb/1/18/Callus 1. jpg/220 px-Callus 1. jpg Picture of GUS reporter: http: //upload. wikimedia. org/wikipedia/commons/thumb/9/9 c/Rice_embryo. png/220 px. Rice_embryo. png http: //books. google. com/books? hl=en&lr=&id=Mx. Fn 0 g. Nq. EWMC&oi=fnd&pg=PA 157&dq=+%22 Gooday%22+%22 Agre ssive+and+defensive+roles+for+chitinases%22&ots=yit. Zd. TSGMH&sig=6 C 0 KXm 93 m. M 3 NLu 2 a 0 R 9 JWIUx_0#v=onepage&q=&f=false Articles: Paul P. Belliveau. "Omalizumab: A Monoclonal Anti-Ig. E Antibody. " Med. Gen. Med. 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. allergies/index. html R. A. Jefferson. The GUS Reporter Gene System. Nature 342, 837 - 838 (14 December 1989). http: //www. nature. com/nature/journal/v 342/n 6251/abs/342837 a 0. 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? dopt=Abstract Website on Latex-Fruit Syndrome: http: //dmd. nihs. go. jp/latex/cross-e. html

Sources continued… Want Milk? Websites Visited: http: //en. citizendium. org/wiki/Bifidobacterium_bifidum http: //www. textbookofbacteriology. net/normalflora. html http: //www. wrongdiagnosis. com/l/lactose_intolerance/prevalence. htm Picture Credits: (2)http: //www. dsm. com/le/en_US/maxilact/html/lactose_intolerance. htm (3)https: //owa. exchange. mit. edu/owa/redir. aspx? C=efb 2 a 8973 caa 4 d 25 a 7203985 c 9 d 18 232&URL=http%3 a%2 f%2 fwww. lactaid. com (1)http: //oregonstate. edu/dept/foodsci/dairy_links. htm (Introduction): http: //badanimalpuns. files. wordpress. com/2009/12/cow. jpg