WHY DON'T GRAVEYARDS FLUORESCE? ANTI-AGING APPLICATIONS OF THE BACTERIAL DEGRADATION OF LYSOSOMAL AGGREGATES





A.D.N.J. de Grey*, J.A.C. Archer

Department of Genetics
University of Cambridge
Downing Street
Cambridge CB2 3EH
UK




The ability to degrade damaged macromolecules is vital for the long-term
survival of any cell. The hydrolytic arsenal present in our lysosomes is
able to break down virtually any such substance present within cells, but a
small proportion is so cross-linked as to be resistant to any endogenous
enzyme. This leads to the accumulation within lysosomes of material that
cannot be degraded, since the only alternative -- exocytosis and excretion
-- is avoided, perhaps due to the risk of toxicity in the extracellular
space. However, eventually the sheer volume of such material compromises
cellular function and integrity. This phenomenon has been neglected by
many biogerontologists, mainly because the substance that has received the
most attention is lipofuscin (the fluorescent lysosomal aggregate most
often found in postmitotic cells), which has not been shown to reach
pathological levels in vivo within a normal lifetime. Other substances
certainly do reach that level, however. Photodamaged rhodopsin
accumulating in the pigmented epithelium of the retina causes macular
degeneration; neurofibrillary tangles and related aggregates are thought to
be causative, not merely diagnostic, of both premature and "normal"
age-related neurodegeneration; and oxidised cholesterol derivatives within
arterial macrophages cause them to degenerate into foam cells, the first --
and a necessary -- stage in the development of atherosclerosis. Hence,
removal of lysosomal aggregates would be an extraordinarily wide-ranging
approach to reversing, not merely retarding, aspects of age-related
cellular and physiological decline.


It might be expected that substances so refractory to lysosomal degradation
would also be very long-lived in the soil after death, and would thus be
detectable in locations such as graveyards. This might not be so, however,
if the soil harbours organisms with hydrolytic capabilities exceeding our
own. Such organisms are indeed present. Within the soil bacteria, the
Nocardioforms (a diverse group of Gram-positive bacteria) play a central
role in the recycling of highly recalcitrant, water-immiscible or poorly
soluble compounds such as aromatic hydrocarbons, halogenated hydrocarbons,
oils, fuels, solvents -- even explosives. If such organisms can break down
highly cross-linked material that is resistant to the human hydrolytic
machinery, it is a conceptually straightforward (though, of course,
technically very challenging) matter to isolate the enzymes responsible and
incorporate them transgenically into affected cells. There should be
little danger to lysosomal integrity, since that is mainly ensured by the
extreme glycosylation of lysosomal membrane proteins and the presence of
highly specialised lipids in the inner leaflet of the lysosomal membrane,
whereas the target material is mainly derived from heterogeneous lipids.
(An even safer mode of delivery may be via engineering of Leishmania, which
naturally targets and survives within macrophage lysosomes.) Preliminary
experiments, whose results will be presented, indicate that such bacteria
can indeed degrade such substances, since they grow on them in the absence
of any other nutrient.




Key words: lipofuscin; foam cells; neurodegeneration; Nocardioforms; gene therapy







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