Presentation for mHealth Israel by Dr. Leon Peshkin, Harvard Medical School, exploring research into Embryology and Longevity, emphasizing Germline and protein aggregates. Includes current overview, anti aging through history, Lifespan Expansion, ALEC: Animal Life Expectancy in Controls, Scalable Platform in Pharmacobiology, Embryos Cleanse Protein Aggregates, Lysosomal switch triggers proteostasis renewal, Xenopus, Rejuvenation Roadmap, Citizen Science Approach to Aging, Xenopus: Maternal Dowry Molecules, Human Disease Modeling

1. Longevity or Radical Life Extension ? First things first.
Leon Peshkin
peshkin @ gmail.com
+33 6 95 33 77 96

2. We are the chosen generation. Singularity is near.
Rejuvenation therapy is almost here. Not one,
several a-la-carte: epigenetic reprogramming, stem
cells, factors from young blood, senolytics,
rapalogues, NAD, etc.
Companies backed up by luminaries from business
and science are rapidly sorting out the remaining
details, helped by the formidable force of Artificial
Intelligence technology known as Deep Learning.
- extend telomeres
- block transposons
- reduce ROS
- clear aggregates
- purge senescent cells
Current view in the field

4. Lifespan Extension ?

6. C57BL/6J mice
ALEC: Animal Life Expectancy in Controls

7. C57BL/6J mice
ALEC: Animal Life Expectancy in Controls

8. Scalable Platform for Pharmacobiology

9. A standardized scalable platform for intervention testing
50

10. 2mm fresh water crustacean
Life expectancy 30d
Parthenogenetic – clonal
Transparent
Large numbers of offspring
Permeable to drugs
Cultivation in small volumes of
water 1/10cc
Normal heart rhythm 200 beat/min
Responds to cardiac drugs
Visible macrophages
Immune system discovered by
Metchnikoff
Eye connected to brain
Complete genome available

11. "Smart tanks" platform
Establish the behavior-based biomarkers of aging using a high-throughput phenotyping platform
and machine learning techniques.
Use these biomarkers in short-term experiments to profile drugs w.r.t. health-span and lifespan.

12. Radical Life Extension – The answer is in Germline

13. Rapid 36 hour development from
egg to organogenesis enables
study disease phenotypes
Source: www.Xenbase.org
Embryos cleanse protein aggregates, why can't we all

14. Why Xenopus
Oocyte maturation is a well-conserved
process in animals. Lysosome activation
occurs during oocyte maturation in C.
elegans (Fig. 2f), but it is not known
whether this is also true of other species.
Therefore, we investigated lysosome
activation during oocyte maturation in
Xenopus laevis. Arrested Xenopus oocytes
had no detectable lysosomal acidity
(100%, n = 9; Fig. 5a). However, acidic
lysosomes were seen in progesterone -
stimulated Xenopus oocytes undergoing
maturation (100%, n = 13; Fig. 5a). Thus,
lysosome acidification during oocyte
maturation is evolutionarily conserved,
and may enhance oocyte proteostasis in
multiple species.
Goudeau, J. & Aguilaniu, H. Carbonylated proteins are eliminated during
reproduction in C. elegans. Aging Cell 9, 991–1003 (2010).

15. are there aggregates ?
which proteins ?
cleansed ?
how ?

16. Why Xenopus ● no one else has a large aged Xenopus colony !
["parabiotic" ovaries]
● homology of oogenesis, progesterone, HCG
(human chorionic gonadotropin)
● "aging ground zero" - thousands of clone embryos
● (yes DNAm clocks, but that's for another day)

17. Why
Xenopus

18. Why Xenopus
Jessus and Ozon, 2004.Biology of the Cell 96.
• A single Xenopus laevis oocyte/egg/zygote contains ~25 ug non-yolk protein (~1000x mouse)
• Previous work by Kenyon suggests a lysosome-mediated protostatic reset in Xenopus oocytes
• Xenopus models have:
• well-annotated reference proteomes and methylation clocks
• established techniques of oocyte/zygote microinjection, gene KD/KO, transgenesis, microdissection, and
transplantation

19. spin out yolk
and organelles,
depolymerize
macromolecules
collect oocytes
and eggs of
different ages
FPLC in highly
denaturing buffer
covalently-crosslinked
protein aggregates
FPLC in native buffer
with salt, heat, detergent
noncovalently-crosslinked
protein aggregates
detect low MW proteins
in high MW fractions by MS
detect low MW proteins
in high MW fractions by MS
Our Strategy
size-exclusion
chromatography

20. ● What changes over time in the germline?
● How do we measure age in the germline and fertilized embryo?
● What are the mechanisms of regulation of rejuvenation ?
● Does it only happen in germ cells and involve meiotic machinery?
● Can you translate and apply what we learn regarding rejuvenation to improve
fertility and/or extend lifespan/rejuvenate somatic tissue to validate importance?
● Frog -> Human, Mouse, ways to screen drugs
Rejuvenation Roadmap

22. THANK YOU
Leon Peshkin
peshkin@gmail.com
06 95 33 77 96

24. Why Xenopus: maternal dowry molecules get early action
Obtain protein synthesis [KS ] and protein degradation [KD ] rates by fitting
a mass-action kinetics equation into the data, also assuming p(t0) = p0 ;
Peshkin L, et al. On the Relationship of Protein and mRNA Dynam
Vertebrate Embryonic Development. Dev Cell. 2015, PMID: 26555

25. Female coelomic cavity (ovary removed)
If the frog is female and has been recently ovulated the oviducts may appear different.
Liver (right
+left lobe)
Fat bodies
lung
oviduct
heart
GI tract
Why Xenopus

26. Xenopus: an excellent system for human disease modeling
Source: www.Xenbase.org, adapted from Wheeler & Brändli 2009 Dev Dyn 238:1287-1308.
• Share 83% human disease genes
• Gene editing: CRISPR KO/KD,
LOF/GOF
• Large eggs and embryos
• Rapid development in vitro
• Cost effective screening
• Genes lost in rodents, present in
amphibia
• Well-defined fate map=> target specific
tissues and organs or left/right side
• Produce hardy eggs year-round
• Frogs are tetrapods
• limbs, fingers & toes [not fins]
• lungs
• 3-chambered heart