RE: Removing lysosomal aggregates; obviating mitochondrial mutations

From: Rafal Smigrodzki (rafal@smigrodzki.org)
Date: Tue Apr 29 2003 - 14:24:25 MDT

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    Aubrey de Grey wrote:

    <snip>

     This seems to me to explain why (as far as we
    can tell) the code disparity is in fact vastly harder for evolution to
    solve than either of the other two hurdles.

    ### We seem to agree that code disparity was the main problem in
    transferring mt genes to the nucleus, although other factors like
    hydrophobicity could play a role as well.

    ------------------------------

    > ### You are talking about the so called "fatty streaks", right? Fatty
    > streaks can fully disappear.

    This is news to me. Can you give references? This certainly flies in
    the face of all I've ever come across on fatty streaks.

    ### See: Desurmont C. Caillaud JM. Emmanuel F. Benoit P. Fruchart JC. Castro
    G. Branellec D. Heard JM. Duverger N. Complete atherosclerosis regression
    after human ApoE gene transfer in ApoE-deficient/nude mice. [Journal
    Article] Arteriosclerosis, Thrombosis & Vascular Biology. 20(2):435-42, 2000
    Feb.

    I also dimly remember reading some Scientific American articles claiming
    reversibility of fatty streaks. I think there were some experiments made on
    the Watanabe rabbits some years ago, involving gene therapy, with regression
    of lesions.

    ----------------------------------------

    > The only way you can tell there is totally indigestible stuff in the
    > plaque is if you cannot digest it in vitro in cell culture

    True. As far as I'm aware, no one has done the simple experiment of
    exposing macrophages to atheroma in culture and seeing what happens.
    Or more likely, it was done 50 years ago and the result was boring...

    > and if injected in vivo in a young animal, it doesn't get phagocytosed.

    I think you meant "OR if injected ..." -- clearly both phagocytosis and
    digestion have to happen, either without the other is useless.

    ### Yes.

    ------------------------------------------
    > ### A different explanation is that there is a dynamic balance between
    > the influx of digestible fatty material, and the ability of endothelium
    > and macrophages to deal with it. As time goes by, the balance tilts
    > towards buildup, with first only occasional macrophages dying, then as
    > the quality of macrophages goes down with age (and it does), there
    > isn't enough to deal with plaque even if it's still digestible, also
    > because of scarring, and calcification.

    I disagree that this is an explanation, because it doesn't explain why
    fatty streaks appear in childhood. Any process of accumulating damage
    (e.g. junk like this) that is already easily detectable in early life
    is by definition primary, not brought on by the misfortune of inhabiting
    the same body as a bunch of other aging tissues (and thus experiencing
    a tilting of a dynamic balance).

    ### Genetically determined obesity (e.g leptin deficiency) manifests early
    in life, and yet we know that maintenance of body weight is a dynamic
    balancing process, and can be tilted early or late in either direction. You
    can have fatty streaks appearing early and developing very fast, if your LDL
    is very high, in hereditary hypercholesterolemia, or they can develop
    slower, if you have only the Western diet going against you. The balance can
    be out of kilter because of hereditary, or acquired changes in your genome,
    or due to non-genetic changes (like formation of insoluble aggregates). So
    far we have insufficient data to answer the question of the specific
    contribution of such factors in PD, AD, and, AFAIK, in most other
    aging-related conditions.

    ------------------------

    > Chinnery and Turnbull did some calculations showing that mtDNA can
    > widely fluctuate and fix mutations into homozygosity if only random
    > processes are taken into account.

    This is one body of work regarding which I tend to have trouble being
    diplomatic. Patrick Chinnery is an excellent medic, but suffice to say
    that we discussed his hypothesis for accumulation by drift five years
    ago, before he'd published anything embarrassing, and he's still having
    trouble understanding the fundamental inconsistency of that model with
    the data, viz. that it predicts that mitochondrial hyperproliferation
    will precede (hence be seen in the absence of) COX-negativity, whereas
    with the sole exception of the MELAS mutation (which seems to have a
    special way to win) the reverse is true, we see abundant COX-negative
    cells that lack any mitochondrial hyperproliferation.

    ### My boss, Davis Parker, likes the idea of clonal expansion, too. Why do
    you think COX negativity must precede hyperproliferation if this hypothesis
    is true?

    ------------------------

    > A mutational ratchet mechanism tends to favor mutated DNA's in
    > a 3 to 1 ratio, but this is not a selective advantage.

    I don't know what underlies your arrival at this ratio, but one can get
    any ratio one likes with suitable assumptions about rates of mutation
    and turnover.

    ### I meant that in the absence of a selective pressure, any sequence tends
    to randomize, and for every non-mutated nucleotide there are three ways of
    mutating it, so you end up with a 3:1 ratio of mutated sequences for each
    base that undergoes mutations.

    -----------------------------------

     The most general argument in favour of selection, though,
    is that we know roughly the mitochondrial half-life (a week to a month
    in rats) and thus we know how many mitochondrial generations it takes
    for cells to be taken over -- and it's very, very few, probably only
    a few times the theoretical minimum of log(2) of the number of mtDNAs
    in the cell. Drift won't do that, whatever the assumptions.

    ### If the effective number of mtDNA genomes undergoing replication is
    smaller than the total number of mitos, then drift can occur pretty fast.

    --------------------------------

    > Actually, once you have mitofection, there are at least two feasible
    > methods to select the mutated mtDNA's out of the cell. Previous
    > attempts failed because of lack of the ability to introduce DNA into
    > mitos, but this has now changed.

    Your turn to lose me. What are you suggesting, in detail?

    ### Well, one is an idea that is undergoing preliminary testing now, the
    other is an idea that evolved in our lab had recently, but was not tested,
    so I am talking about highly speculative issues. I really can't discuss the
    details.

    Rafal



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