HDAC Article: - Trehalose: Sweet Rescue
Trehalose: Sweet Rescue
-- LaVonne Veatch Goodman, M.D.     print-friendly ]
Cumberland, the makers of Sweet & Low, is now packaging trehalose in 3 lb containers that can be obtained over the web http://www.trehalose.biz/ . This is a very "clean" product --FDA "GRAS" approved as a safe food.

Trehalose is a food product which hasn't been available retail in the US before. The dose we use in HDDW trials is 25 gms three times a day. The price they quote means about $1/day. Though our numbers are small, the trend in our trials suggests that trehalose is helpful at all stages of HD, but the earlier the better.

If you use this I recommend you start with 10 grams a dose and work your way up. Unfortunately about 25% of HDDW participants had diarrhea. I think it is ok, even good, in diabetics. Its use resulted in better control of sugars in one of my Huntington's patients.


-- LaVonne Veatch Goodman, M.D.

 

Trehalose is a naturally occurring disaccharide with known protein and membrane stabilizing capability. Because of these unique chemical properties, this molecule has been the focus of study in several neurodegenerative diseases which are associated with the misfolding of disease-specific proteins. These conditions include Alzheimer?s disease (AD) an amyloid proteinopathy, Huntington?s disease (HD), an expanded polyglutamine proteinopathy, and oculopharyngeal muscular dystrophy (OPMD), an expanded polyalanine proteinopathy.

In each disease, specific misfolded aggregate-prone proteins are resistant to the normal cellular processes of protein turnover and accumulate in insoluble inclusions in regions specific to each disease. While insoluble aggregates correlate with disease progression, there is increasing evidence that the initiating and most toxic events are caused by soluble protein oligimers or microaggregates. Trehalose is thought to work by interfering with production or enhancing destruction of toxic fragments.

Natural Functions of Trehalose

One of the fascinating aspects of trehalose is its presence in various organisms that can survive at the extremes of temperature and dehydration. This observation led to work which showed that trehalose is a naturally occurring reducer of cell stress, protecting these organisms from extremes in heat shock and osmotic stress (Crowe 2002). Trehalose is thought to act by altering or replacing the water shell that surrounds lipid and protein macromolecules (Colaco et al. 1995). It is thought that its flexible glycosidic bond allows trehalose to conform with the irregular polar groups of marcromolecules. In so doing, it is able to maintain the three-dimensional structure of these biologic molecules under stress, preserving biologic function.

Therapeutic Uses

As an extension of its natural capability to protect biological structures, trehalose has been used for the preservation and protection of biologic materials. It stabilizes bioactive soluble proteins such as monoclonal antibodies and enzymes for medical use (Colaco et al. 1992). It stabilizes proteins for inhaled use (Strickley and Anderson 1997). It is used to preserve cellular blood products for transfusion and greatly extends the shelf life of platelets (Crowe et al. 2003), and cord blood (Zhang et al. 2003). It is used to preserve embryos during freeze-drying where it increases viability (Suzuki et al. 1996). It is used in cryopreservation of transplant cells and tissue where it has been shown to increase viability (Beattie et al. 1997) and decrease host immune response (Erdag et al. 2002).

Building on extensive study in multiple biologic systems that describe its ability to inhibit lipid and protein misfolding (Singer and Lindquist 1998), trehalose has become an attractive molecule for study in neurodegenerative disease characterized by protein misfolding and aggregate pathology. Such diseases include Alzheimer?s and Parkinson?s disease, and the less common triplet repeat diseases. Recent scientific publications describe trehalose benefit in model systems that recapitulate aggregate pathology that characterize Alzheimer?s (AD), Huntington?s (HD), and occulopharyngeal muscular dystrophy (OPMD).

Huntington?s Disease

Trehalose benefit was first shown in Huntington?s model systems (Tanaka et al. 2004). Huntington?s is an autosomal dominant neurodegenerative disease which presents with cognitive impairment, involuntary choreiform movements, and psychiatric manifestations. Onset is generally in midlife, but can occur in childhood and old age. It inexorably progresses to disability and death over a 10-25 year period. Huntington?s is characterized by an expanded CAG repeat within the first exon of the huntington gene. The mutant protein generated has an expanded polyglutamine (polyGN) tract. The pathologic hallmark of this, and other polyGN diseases is the formation of aggregates, containing misfolded mutant protein in both cytoplasm and nucleus of affected cells.

Tanaka, et al, demonstrated that trehalose inhibits polyglutamine-mediated protein aggregation of a model polyGN protein in vitro solution, and that it decreases aggregate formation and prolongs viability in a model cell culture. This same group went on to show that trehalose ameliorated motor symptoms, decreased aggregate number and size, and prolonged life by 20% in the R6/2 transgenic mouse model of Huntington?s.

In Vitro: Because huntingtin protein is not soluble, the investigators studied the effects of trehalose on myoglobin mutant proteins Gln35 (35 glutamine repeats) and MbGln 12 (12 repeats). The Gln35 molecules mimic the extended polyglutamine stretches of mutant huntingtin protein, the Gln12 corresponds to the normal huntingtin protein. Using spectroscopic means, the authors showed that trehalose interacts with the expanded polyglutamine (above 12 repeats) tract of the proteins, stabilizing structure, and inhibiting protein aggregation at the initial stage of aggregate formation.

Cell Culture: They next studied the effects of trehalose in the Neuro2a cell model that expresses a truncated N-terminal huntingtin molecule containing an expanded polyglutamine stretch of 60 or 150 glutamine repeats. They found that extracellular trehalose application decreased intracellular aggregates and improved cell viability in a dose-dependent fashion. Intracellular trehalose concentration was not reported. In a related experiment, Neuro2a cells were genetically engineered to transiently produce intracellular trehalose. These cells had measured levels of trehalose (ll.8 +/- 1.5 nmol/ug protein) which was associated with decreased intracellular huntingtin aggregates and increased cell viability more than 50% over baseline.

Mammalian in vivo model: They next studied the R6/2 mouse model that had received 2% trehalose in drinking water. They determined that trehalose (0.11 +/- O.O2 nmol/ug protein) was present in brain homogenates of treated mice. They further showed that the trehalose-treated mice had reduced brain atrophy, and decreased the number of intranuclear aggregates by 55% from that observed in control mice. Further, the treated mice demonstrated improved motor function and life extension by 20%.

Further evidence of brain neuronal penetration is provided by a report from the laboratory of H. Nguyen and O. Riess from the University of Tuebingen (Nguyen et al. 2005). In data presented at the 2005 Society for Neuroscience meetings, they reported that 2% oral administration of trehalose is sufficient to favorably shift gene expression of a rat transgenic HD model.

Trehalose was among the first molecules chosen for review from the SET-HD list compiled at the National Institute for Neurologic Disease and Stroke (NINDS). Based on Tanaka?s work, trehalose is now a lead compound for use in people with Huntington?s and has potential for delaying onset or decreasing progression rate in this disease

Oculopharyngeal Muscular Dystrophy

Oculopharyngeal muscular dystrophy (OPMD) is an autosomal dominant disease that presents in the fifth or sixth decade of life with dysphagia, ptosis and proximal limb weakness. It is caused by an abnormal expansion of a polyalanine tract within the coding region of poly-A binding protein nuclear 1 (PAβPN1). This mutant protein demonstrates cellular pathology similar to Huntington?s in that it forms aggregates. In OPMD, the culprit mutant protein forms aggregates in nuclei of skeletal muscle fibers, causing injury which culminates in cell dysfunction and death.

Trehalose has been found beneficial in transgenic model systems for this disease (Davies et al. 2006). The authors have shown that trehalose reduces aggregate formation and prolongs viability in the cell model of this disease, and that it delays onset, ameliorates symptoms, and prolongs life in the mouse model of this disease.

Cell Culture: Using a COS-7 cell line transfected with plasmids encoding for 17 alanine repeats, they showed that application of trehalose to the culture medium decreased aggregates by almost 50%, and increased cell viability by 30%.

Mammalian in vivo model: They next studied the A17-1 mouse model of OPMD. Trehalose supplied in a dose of 2% in drinking water significantly delayed disease onset, maintained muscle strength, and significantly decreased aggregate formation and cell death.

The authors summarize, that based on their work, trehalose has potential for delaying onset and decreasing progression rate in people at risk for this disease.

Alzheimer?s Disease

Sporadic Alzheimer?s disease is pathologically characterized by aggregation of small beta-amyloid (Aβ) peptides into amyloid plaques and neurofibrillary tangles. As in other proteinopathy neurodegenerative disease, Alzheimer?s progression is correlated with increasing aggregate load of amyloid, though toxicity is increasingly linked to the formation of oligomeric forms of Aβ peptides (Walsh et al. 2005).

Trehalose was studied in vitro, and in cell models of Alzheimer?s by using the proteins beta-amyloid 40 and 42, which are the proteins most prominently implicated in Alzheimer?s (Liu et al. 2005). Although Aβ40 is the most prominent peptide (10:1), Aβ42 is more toxic. The authors report effects of trehalose on aggregation in vitro, and neurotoxicity in a cell model, using solutions of Aβ peptide either singly or in combination.

In Vitro Aggregation Formation Studies; Aβ40 In vitro: In solution at physiologic concentration, Aβ40, incubated alone, displays a time dependent increase in spontaneous aggregation. Co-incubation with trehalose inhibits aggregation of this peptide in a dose-dependent manner. High concentration trehalose completely inhibited aggregation of this peptide.

Aβ42: In vitro: In physiologic concentrations, Aβ42, when incubated alone aggregates more quickly than Aβ40. Aβ42 has an additional two hydrophobic amino acids, making this peptide more aggregate prone than Aβ40. While trehalose inhibited aggregation of Aβ42 in a dose-dependent manner, it was significantly less efficient than the corresponding Aβ40. High dose trehalose prevented only 50% aggregation of this more toxic peptide.

Combination: They also reported that trehalose inhibits aggregation of the two peptides in solution of a physiological ratio (10:1) of combined Aβ40 and 42 monomers.

In Vitro Aggregate Morphology Studies: They next examined characteristics of Aβ aggregate morphology when incubated with trehalose.

Aβ40: In addition to aggregates, the Aβ40 control sample showed the presence of small oligomers and many thin protofibrils. When Aβ40 was co-incubated with trehalose, the number of filaments and oligomers substantially decreased in a dose dependent manner.

Aβ42: Likewise, the control Aβ42 aggregate sample showed many fibril formations and the presence of small oligomers. However, in this case, trehalose coincubation did not significantly reduce oligomer or fibril concentration.

There was no reported study on trehalose effect using a combination of peptides.

Cell Model

The authors used a SH-SY5Y model neuron cell system for study of Aβ toxicity. Co-incubation of these cells with either Aβ40 or Aβ42 resulted in cell toxicity in a dose dependent manner. Aβ40: When incubated alone, Aβ40 exhibited substantial cytotoxicity to SH-SY5Y cells. However, co-incubation with trehalose showed a dose-dependent decrease in toxicity, with nearly complete protection against Aβ40 toxicity at high dose. Aβ42: Whether incubated alone or with trehalose, Aβ42 showed similar toxicity to SH-Y5Y cells. There was no protective effect at any dose. There was no reported study of trehalose effect using a combination of peptides in cell medium. The authors summarize that based on their work, trehalose has potential as part of a therapeutic ?cocktail? strategy for treating Alzheimer?s because of its effect on Aβ40, the predominant form of Aβ peptide in amyloid plaques. They add that complementary methods for controlling the more toxic Aβ42 oligomerization would be necessary.

Summary

Although trehalose absorption in humans has not been well studied, a small fraction (0.5%) is likely to be absorbed by passive diffusion, as has been demonstrated for other dissacharides (van Elburg et al. 1995). In mammalian cell culture, trehalose is moved from the extracellular to intracellular compartment via a fluid phase endocytotic mechanism, and is dependent on extracellular concentration (Oliver et al. 2004). Trehalose dose measured in brain homogenates of mammalian models is quite low, but Tanaka, et al postulate that low concentration may exert effect because trehalose is resistant to hydrolysis. In cell culture, once introduced, intracellular trehalose concentration does not change for extended periods of time (Eroglu et al. 2000). Accordingly, its structure/activity benefits would be expected to persist for relatively extended times. If planned studies demonstrate brain or cerebral spinal fluid absorption, trehalose will open a new avenue of potential therapy for the prevention and treatment of multiple neurodegenerative diseases. This review summarized evidence for protective benefit in models of Huntingtin?s disease, oculopharyngeal muscular dystrophy, and Alzheimer?s. Although not studied, Parkinson?s disease and amyotrophic lateral sclerosis display aggregate pathology that may be amenable to similar response.

Taken together, these studies suggest that ingestion of trehalose, a naturally occurring disaccharide, may have protective effect in promoting brain health. There is potential for both prevention and treatment of neurodegenerative diseases. This is especially exciting in light of this agent?s well documented safety profile (Richards et al. 2002) and GRAS approval for use in the United States.

References

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- published -0-4--2006