Acetaldehyde Detoxification Research Articles

Aldehyde dehydrogenase 1 family: A potential molecule target for diseases.

Aldehyde dehydrogenase 1 (ALDH1), a crucial aldehyde metabolizing enzyme, has six family members. The ALDH1 family is expressed in various tissues, with a significant presence in the liver. It plays a momentous role in several pathophysiological processes, including aldehyde detoxification, oxidative stress, and lipid peroxidation. Acetaldehyde detoxification is the fundamental function of the ALDH1 family in participating in vital pathological mechanisms. The ALDH1 family can catalyze retinal to retinoic acid (RA) that is a hormone-signaling molecule and plays a vital role in the development and adult tissues. Furthermore, there is a need for further and broader research on the role of the ALDH1 family as a signaling molecule. The ALDH1 family is widely recognized as a cancer stem cell (CSC) marker and plays a significant role in the proliferation, invasion, metastasis, prognosis, and drug resistance of cancer. The ALDH1 family also participates in other human diseases, such as neurodegenerative diseases, osteoarthritis, diabetes, and atherosclerosis. It can inhibit disease progression by inhibiting/promoting the expression/activity of the ALDH1 family. In this review, we comprehensively analyze the tissue distribution, and functions of the ALDH1 family. Additionally, we review the involvement of the ALDH1 family in diseases, focusing on the underlying pathological mechanisms and briefly talk about the current status and development of ALDH1 family inhibitors. The ALDH1 family presents new possibilities for treating diseases, with both its upstream and downstream pathways serving as promising targets for therapeutic intervention. This offers fresh perspectives for drug development in the field of disease research.

Aged Garlic Extract Prevents Alcohol-Induced Cytotoxicity through Induction of Aldehyde Dehydrogenase 2 intheLiverofMice.

Acetaldehyde is a highly toxic primary metabolite of ethanol, and converts to nontoxic acetic acid by aldehyde dehydrogenase (ALDH). Accumulation of acetaldehyde causes significant damage to human body. Aged garlic extract (AGE) is a functional food material and possesses various health beneficial effects. This study investigates whether AGE contributes to acetaldehyde detoxification through ALDH induction and its underlying mechanism. C57BL/6J mice are orally administrated 10-1000mgkg-1 body weight (BW) of AGE for 1week before ethanol administration. AGE suppresses ethanol-caused accumulation of acetaldehyde level in the plasma through inducing mitochondrial ALDH2 but not cytosolic ALDH1A1. AGE also induces antioxidant enzymes, heme oxygenase-1, and NAD(P)H:quinone oxidoreductase 1, resulting in prevention of lipid peroxidation in the liver. In HepG2 cells, AGE prevents ethanol- and acetaldehyde-caused cytotoxicity. AGE induces mitochondrial ALDH2 through activating nuclear factor-erythroid 2-related factor 2 (Nrf2). AGE inhibits protein degradation of Nrf2 and enhances protein degradation of kelch-like ECH-associated protein 1. Furthermore, S-allyl cysteine and S-allyl mercaptocysteine as the bioactive compounds in AGE also induce ALDH2 and Nrf2. AGE prevents acetaldehyde-induced hepatotoxicity through enhancing acetaldehyde detoxification through Nrf2-dependent induction of mitochondrial ALDH2.

The role of aldehyde dehydrogenase 2 in cardiovascular disease.

Aldehyde dehydrogenase 2 (ALDH2) is a mitochondrial enzyme involved in the detoxification of alcohol-derived acetaldehyde and endogenous aldehydes. The inactivating ALDH2 rs671 polymorphism, present in up to 8% of the global population and in up to 50% of the East Asian population, is associated with increased risk of cardiovascular conditions such as coronary artery disease, alcohol-induced cardiac dysfunction, pulmonary arterial hypertension, heart failure and drug-induced cardiotoxicity. Although numerous studies have attributed an accumulation of aldehydes (secondary to alcohol consumption, ischaemia or elevated oxidative stress) to an increased risk of cardiovascular disease (CVD), this accumulation alone does not explain the emerging protective role of ALDH2 rs671 against ageing-related cardiac dysfunction and the development of aortic aneurysm or dissection. ALDH2 can also modulate risk factors associated with atherosclerosis, such as cholesterol biosynthesis and HDL biogenesis in hepatocytes and foam cell formation and efferocytosis in macrophages, via non-enzymatic pathways. In this Review, we summarize the basic biology and the clinical relevance of the enzymatic and non-enzymatic, tissue-specific roles of ALDH2 in CVD, and discuss the future directions in the research and development of therapeutic strategies targeting ALDH2. A thorough understanding of the complex roles of ALDH2 in CVD will improve the diagnosis, management and prognosis of patients with CVD whoharbour the ALDH2 rs671 polymorphism.

EP16.04-007 ALDH2 Downregulation, Aberrant DNA Methylation, and Loss of Stemness Pathways as an Enhanced Biomarker in Lung Adenocarcinoma

Lung adenocarcinoma (LUAD) is the most common type of lung cancer and is one of the major causes of growing mortality. Previous studies showed an association between LUAD and Aldehyde Dehydrogenase 2 (ALDH2), an essential enzyme involved in the detoxification of acetaldehyde. We aimed at discovering more informative prognosis signatures for early LUAD detection and targets for LUAD treatment by investigating the expression level, epigenetic mechanism, and signaling activities of ALDH2 in LUAD patients.

The seed biochemical signature as a potent marker for water logging tolerance in maize

Plant breeders are in the need for convenient, reproducible, reliable and rapid screening methods to be used as a proxy for water logging tolerance for a large number of genotypes. Addressing this need, the present study was carried in order to identify potential biochemical markers in the seeds of 10 contrasting maize (Zea mays L.) genotypes differing in their tolerance against water logging. Results clearly indicate that tolerant genotypes inherently exhibit better antioxidant system as evident by higher activities of catalase and glutathione reductase than the susceptible genotypes hence minimizes resulting oxidative stress. Significantly higher activities of anaerobic metabolism enzymes viz. alcohol dehydrogenase, aldehyde dehydrogenase and pyruvate decarboxylase in tolerant maize genotypes signifies efficient system for the recycling of NADH for the continuation of glycolysis and simultaneous detoxification of acetaldehyde thus provides prolonged survival of tolerant plants under water logging. Higher Sucrose synthase (SuSy) activity in tolerant genotypes over the susceptible genotypes indicates uninterrupted supply of reducing sugars for glycolytic activity. The content of total and reducing sugars content was found 27% higher in tolerant genotypes, which is in agreement with the SuSy activity. Soluble sugars analysis through reverse phase HPLC revealed higher relative concentration of fructose and glucose in tolerant maize genotypes than susceptible genotypes. Tolerant genotypes also exhibited significantly lower phytic acid and higher proline content. Hence the observed biochemical diversity could be useful for screening and development of water logging stress-tolerant genotypes of maize.

Aldehyde dehydrogenase 2 protects against acute kidney injury by regulating autophagy via Beclin-1 pathway

The mitochondrial enzyme aldehyde dehydrogenase 2 (ALDH2) catalyzes the detoxification of acetaldehyde and endogenous lipid aldehydes. Approximately 40% of East Asians, accounting for 8% of the human population, carry the E504K mutation in ALDH2 that leads to accumulation of toxic reactive aldehydes and increases the risk for cardiovascular disease, cancer, and Alzheimer disease, among others. However, the role of ALDH2 in acute kidney injury (AKI) remains poorly defined and is therefore the subject of the present study using various cellular and organismal sources. In murine models, in which AKI was induced by either the contrast agent iohexol or renal ischemia/reperfusion, KO, activation/overexpression of ALDH2 were associated with increased and decreased renal injury, respectively. In murine renal tubular epithelial cells (RTECs), ALDH2 upregulated Beclin-1 expression, promoted autophagy activation, and eliminated ROS. In vivo and in vitro, both 3-MA and Beclin-1 siRNAs inhibited autophagy and abolished ALDH2-mediated renoprotection. In mice with iohexol-induced AKI, ALDH2 knockdown in RTECs using AAV-shRNA impaired autophagy activation and aggravated renal injury. In human renal proximal tubular epithelial HK-2 cells exposed to iohexol, ALDH2 activation potentiated autophagy and attenuated apoptosis. In mice with AKI induced by renal ischemia/reperfusion, ALDH2 overexpression or pretreatment regulated autophagy mitigating apoptosis of RTECs and renal injury. In summary, our data collectively substantiate a critical role of ALDH2 in AKI via autophagy activation involving the Beclin-1 pathway.

Enhancement of Solubility, Purification, and Inclusion Body Refolding of Active Human Mitochondrial Aldehyde Dehydrogenase 2.

Mitochondrial aldehyde dehydrogenase 2 (ALDH2) is predominantly linked with acetaldehyde detoxification in the second stage of alcohol metabolism. To intensively study ALDH2 function, a higher purity and uniform composition of the protein is required. An efficient Escherichia coli system for ALDH2 expression was developed by using His and a small ubiquitin-related modifier fusion tag. Most of the recombinant ALDH2s were expressed in the form of inclusion bodies. The ALDH2-enriched inclusion bodies were denatured with 6 M guanidine hydrochloride, and then ALDH2 was ultrafitrated. Finally, ALDH2 was successfully purified through affinity and gel filtration chromatography. The purified ALDH2 was finally preserved by the vacuum freeze-drying method, and its purity was determined to be higher than 95%, with a final media yield of 33.89 mg/L. The specific activity of ALDH2 was 6.1 × 104 U/mg. This work was the first to report pET-SUMO-ALDH2 recombinant plasmid expression in Escherichia coli, and the inclusion bodies were isolated and refolded. Finally, the purified ALDH2 had relatively higher purity, yield, and biological activity.

Intake of Lactobacillus rhamnosus GG (LGG) fermented milk before drinking alcohol reduces acetaldehyde levels and duration of flushing in drinkers with wild-type and heterozygous mutant ALDH2: a randomized, blinded crossover controlled trial.

Alcohol consumption leads to acetaldehyde accumulation, especially in people with mutant aldehyde dehydrogenase 2 gene (ALDH2). Novel strategies to promote acetaldehyde detoxification are required to prevent alcohol-related toxicity. Probiotic bacteria such as Lactobacillus rhamnosus GG (LGG) were shown to have in vitro capacity to detoxify acetaldehyde. This randomized, blinded, placebo-controlled cross-over trial investigated the effect of LGG fermented milk in people with ALDH2 polymorphisms after moderate alcohol intake. Ten healthy wild-type and ten heterozygous mutant ALDH2 Thai men were block randomized into two groups. Each group consumed a different sequence of 150 mL fermented milk containing 108 CFU mL-1 LGG and lactic-acidified milk (placebo), followed by five glasses of beer (0.4 g ethanol per kg body weight), with a one-week wash-out. Consuming LGG fermented milk before alcohol reduced areas under the response curves of blood and salivary acetaldehyde in wild-type and heterozygous mutant ALDH2 individuals (p < 0.05 and p < 0.01, respectively). Interestingly, participants with mutant ALDH2 responded better than wild-type participants for salivary acetaldehyde (90% vs. 70%, p < 0.001). Their durations of flushing were reduced when consuming LGG milk. Regardless of ALDH2 status, 105 CFU mL-1 LGG was retained in saliva at least 3.5 h after milk consumption. In conclusion, intake of LGG fermented milk before drinking alcohol reduces blood and salivary acetaldehyde levels and duration of flushing in drinkers with wild-type and heterozygous mutant ALDH2. The addition of exogenous capacity to detoxify acetaldehyde using the probiotic product could be a potential strategy to promote the alleviation of exposure to reactive and carcinogenic acetaldehyde associated with alcohol drinking in individuals with defective ALDH2 enzyme.

Alcohol-derived DNA crosslinks are repaired by two distinct mechanisms.

Acetaldehyde is a highly reactive, DNA-damaging metabolite that is produced upon alcohol consumption1. Impaired detoxification of acetaldehyde is common in the Asian population, and is associated with alcohol-related cancers1,2. Cells are protected against acetaldehyde-induced damage by DNA crosslink repair, which when impaired causes Fanconi anaemia (FA), a disease resulting in failure to produce blood cells and a predisposition to cancer3,4. The combined inactivation of acetaldehyde detoxification and the FA pathway induces mutation, accelerates malignancies and causes the rapid attrition of blood stem cells5-7. However, the nature of the DNA damage induced by acetaldehyde and how this is repaired remains a key question. Here we generate acetaldehyde-induced DNA interstrand crosslinks and determine their repair mechanism in Xenopus egg extracts. We find that two replication-coupled pathways repair these lesions. The first is the FA pathway, which operates using excision-analogous to the mechanism used to repair the interstrand crosslinks caused by the chemotherapeutic agent cisplatin. However, the repair of acetaldehyde-induced crosslinks results in increased mutation frequency and an altered mutational spectrum compared with the repair of cisplatin-induced crosslinks. The second repair mechanism requires replication fork convergence, but does not involve DNA incisions-instead the acetaldehyde crosslink itself is broken. The Y-family DNA polymerase REV1 completes repair of the crosslink, culminating in a distinct mutational spectrum. These results define the repair pathways of DNA interstrand crosslinks caused by an endogenous and alcohol-derived metabolite, and identify an excision-independent mechanism.

Acetaldehyde production by Rothia mucilaginosa isolates from patients with oral leukoplakia

ABSTRACT Rothia mucilaginosa has been found at high abundance on oral leukoplakia (OLK). The ability of clinical isolates to produce acetaldehyde (ACH) from ethanol has not been investigated. The objective of the current study was to determine the capacity of R. mucilaginosa isolates recovered from OLK to generate ACH. Analysis of R. mucilaginosa genomes (n = 70) shows that this species does not normally encode acetaldehyde dehydrogenase (ALDH) required for detoxification of ACH. The predicted OLK metagenome also exhibited reduced ALDH coding capacity. We analysed ACH production in 8 isolates of R. mucilaginosa and showed that this species is capable of generating ACH in the presence of ethanol. The levels of ACH produced (mean = 53 µM) were comparable to those produced by Neisseria mucosa and Candida albicans in parallel assays. These levels were demonstrated to induce oxidative stress in cultured oral keratinocytes. This study shows that R. mucilaginosa can generate ACH from ethanol in vitro at levels which can induce oxidative stress. This organism likely contributes to oral ACH levels following alcohol consumption and the significance of the increased abundance of R. mucilaginosa in patients with potentially malignant disorders requires further investigation.

YKL107W from Saccharomyces cerevisiae encodes a novel aldehyde reductase for detoxification of acetaldehyde, glycolaldehyde, and furfural.

The aldehyde reductases from the short-chain dehydrogenase/reductase (SDR) family were identified as a series of critical enzymes for the improved tolerance of Saccharomyces cerevisiae to the aldehydes by catalyzing the detoxification reactions of aldehydes. Herein, we report that a novel aldehyde reductase Ykl107wp deduced from YKL107W from S. cerevisiae belongs to the classical SDR group and can catalyze the reduction reactions of acetaldehyde (AA), glycolaldehyde (GA), furfural (FF), formaldehyde (FA), and propionaldehyde (PA) but cannot reduce the six representative ketones. Ykl107wp displayed the best maximum velocity (Vmax), catalytic rate constant (Kcat), catalytic efficiency (Kcat/Km), and highest affinity (Km) to acetaldehyde. The optimum pH of Ykl107wp was 6.0 for the reduction of AA and 7.0 for the reduction of GA and FF, and the optimum temperatures were 40, 35, and 30°C for the reduction of AA, GA, and FF, respectively. Ykl107wp for the reduction of AA was greatly affected by metal ions, chemical additives, and salts and showed poor thermal and pH stability, but its stability was slightly affected by a substrate. Ykl107wp was localized in endoplasmic reticulum and prevented the yeast cells from damage caused by furfural via the detoxification of furfural to furfural alcohol. This research provides guidelines for the study of uncharacterized classical SDR aldehyde reductases and exploration of their protective mechanisms on the corresponding organelles.

Targeting ALDH2 in Atherosclerosis: Molecular Mechanisms and Therapeutic Opportunities.

Aldehyde dehydrogenase 2 (ALDH2) is an important member of the functional aldehyde dehydrogenases (ALDHs) family in human beings, playing a fundamental role in the detoxification of acetaldehyde and other aldehydes. In recent years, a number of researches have given attention to the association between ALDH2 and atherosclerosis, which provided insights on targeting ALDH2 for therapeutic intervention of atherosclerosis. In this review, these inspiring studies will be discussed, and the clinical implications and concerns will be expounded.

Ethanol Dehydrogenase I Contributes to Growth and Sporulation Under Low Oxygen Condition via Detoxification of Acetaldehyde in Metarhizium acridum

The entomopathogenic fungi encounter hypoxic conditions in both nature and artificial culture. Alcohol dehydrogenases (ADHs) are a group of oxidoreductases that occur in many organisms. Here we demonstrate that an alcohol dehydrogenase I, MaADH1, in the locust-specific fungal pathogen, Metarhizium acridum, functions in acetaldehyde detoxification mechanism under hypoxic conditions in growth and sporulation. The MaADH1 was highly expressed in sporulation stage under hypoxic conditions. Compared with a wild-type strain, the ΔMaADH1 mutant showed inhibited growth and sporulation under hypoxic conditions, but no impairment under normal conditions. Under hypoxic conditions, ΔMaADH1 mutant produced significant decreased alcohol, but significant increased acetaldehyde compared to wild type. M. acridum was sensitive to exogenous acetaldehyde, exhibiting an inhibited growth and sporulation with acetaldehyde added in the medium. MaADH1 did not affect virulence. Our results indicated that the MaADH1 was critical to growth and sporulation under hypoxic stress by detoxification of acetaldehyde in M. acridum.

Heterologous Expression of Aldehyde Dehydrogenase in Lactococcus lactis for Acetaldehyde Detoxification at Low pH.

Aldehyde dehydrogenase (E.C. 1.2.1.x) can catalyze detoxification of acetaldehydes. A novel acetaldehyde dehydrogenase (istALDH) from the non-Saccharomyces yeast Issatchenkia terricola strain XJ-2 has been previously characterized. In this work, Lactococcus lactis with the NIsin Controlled Expression (NICE) System was applied to express the aldehyde dehydrogenase gene (istALDH) in order to catalyze oxidation of acetaldehyde at low pH. A recombinant L. lactis NZ3900 was obtained and applied for the detoxification of acetaldehyde as whole-cell biocatalysts. The activity of IstALDH in L. lactis NZ3900 (pNZ8148-istALDH) reached 36.4UmL-1 when the recombinant cells were induced with 50ngmL-1 nisin at 20°C for 2h. The IstALDH activity of recombinant L. lactis cells showed higher stability at 37°C and pH 4.0 compared with the crude enzyme. L. lactis NZ3900 (pNZ8148-istALDH) could convert acetaldehyde at pH 2.0 while the crude enzyme could not. Moreover, the resting cells of L. lactis NZ3900 (pNZ8148-istALDH) showed a 2.5-fold higher activity and better stability in catalyzing oxidation of acetaldehyde at pH 2.0 compared with that of Escherichia coli expressing the IstALDH. Taken together, the L. lactis cells expressing recombinant IstALDH are potential whole-cell biocatalysts that can be applied in the detoxification of aldehydes.

BRCA1 and BRCA2 tumor suppressors protect against endogenous acetaldehyde toxicity

Maintenance of genome integrity requires the functional interplay between Fanconi anemia (FA) and homologous recombination (HR) repair pathways. Endogenous acetaldehyde, a product of cellular metabolism, is a potent source of DNA damage, particularly toxic to cells and mice lacking the FA protein FANCD2. Here, we investigate whether HR‐compromised cells are sensitive to acetaldehyde, similarly to FANCD2‐deficient cells. We demonstrate that inactivation of HR factors BRCA1, BRCA2, or RAD51 hypersensitizes cells to acetaldehyde treatment, in spite of the FA pathway being functional. Aldehyde dehydrogenases (ALDHs) play key roles in endogenous acetaldehyde detoxification, and their chemical inhibition leads to cellular acetaldehyde accumulation. We find that disulfiram (Antabuse), an ALDH2 inhibitor in widespread clinical use for the treatment of alcoholism, selectively eliminates BRCA1/2‐deficient cells. Consistently, Aldh2 gene inactivation suppresses proliferation of HR‐deficient mouse embryonic fibroblasts (MEFs) and human fibroblasts. Hypersensitivity of cells lacking BRCA2 to acetaldehyde stems from accumulation of toxic replication‐associated DNA damage, leading to checkpoint activation, G2/M arrest, and cell death. Acetaldehyde‐arrested replication forks require BRCA2 and FANCD2 for protection against MRE11‐dependent degradation. Importantly, acetaldehyde specifically inhibits in vivo the growth of BRCA1/2‐deficient tumors and ex vivo in patient‐derived tumor xenograft cells (PDTCs), including those that are resistant to poly (ADP‐ribose) polymerase (PARP) inhibitors. The work presented here therefore identifies acetaldehyde metabolism as a potential therapeutic target for the selective elimination of BRCA1/2‐deficient cells and tumors.

Abstract 3280: Associations between ALDH1A1 polymorphisms, alcohol consumption, and mortality among Hispanic and non-Hispanic White women diagnosed with breast cancer: The Breast Cancer Health Disparities Study

Abstract ALDH1A1 is a marker of normal tissue stem cells and cancer stem cells, where it is involved in self-renewal, differentiation and self-protection. ALDH1A1 is also associated with alcohol sensitivity and dependence, and functions in the detoxification of acetaldehyde. Few epidemiological studies have examined the association between ALDH1A1 and mortality among women with breast cancer (BC). To date, none of these studies have included Hispanic women or evaluated the modifying effect of alcohol consumption. Using data from the Breast Cancer Health Disparities Study, we evaluated the associations between nine SNPs of ALDH1A1 (rs348481, rs168351, rs1888202, rs63319, rs722921, rs348461, rs348463, rs7027604, rs1424482) and overall mortality among 1134 Hispanic and 1160 non-Hispanic white (NHW) women diagnosed with incident invasive BC. Demographic and lifestyle factors were collected via in-person interviews. Modes of inheritance were considered for each SNP accordingly using their genotypes. Multivariate Cox proportional hazard regression models were used to calculate hazard ratios (HRs) and 95% confidence interval (CIs). Models were adjusted for percentage of Native American (NA) ancestry, BC summary stage, and study site. To detect effect modification, interaction terms were created between percentage of NA ancestry (low ≤0.28 versus high &amp;gt;0.28), alcohol consumption (ever versus never) and the SNPs. The likelihood ratio test was used to evaluate whether the interaction terms were significant. The Bonferroni correction was used to adjust for multiple comparisons. After a median follow-up time of 11 years from BC diagnosis to death, a total of 542 deaths occurred. The following four SNPs were significantly associated with overall mortality: rs1424482 (HRCC =1.33; 95% CI 1.04-1.70), rs63319 (HRAA =1.30; 95% CI 1.06-1.60), rs7027604 (HRAA =1.37; 95% CI 1.11-1.70), and rs722921 (HRTA/AA =0.78; 95% CI 0.64-0.96). However, only rs7027604 remained significant after adjustment for multiple comparisons (Padj=0.035). Among women with low NA ancestry, rs63319 significantly increased risk of mortality (HRAA= 1.55; 95% CI 1.20-1.99), while a non-significant inverse association was observed among women of high NA ancestry. Among ever drinkers, rs1888202 significantly decreased risk of mortality (HRCG/GG=0.62; 95%CI 0.45-0.85), while a non-significant inverse association was observed among non-drinkers. The adjusted P-values for these interactions were not significant. This study provides evidence that rs7027604 is associated with worse prognosis after BC diagnosis among Hispanic and NHW women. Future BC survival studies examining the relationship between ALDH1A1 and mortality should also explore the modifying effects of alcohol consumption with rs1888202 and NA ancestry with rs63319. Citation Format: Zhiyu Xia, Avonne E. Connor, Kathy B. Baumgartner, Richard N. Baumgartner, Stephanie D. Boone, Lisa M. Hines, Esther M. John, Roger K. Wolff, Martha L. Slattery. Associations between ALDH1A1 polymorphisms, alcohol consumption, and mortality among Hispanic and non-Hispanic White women diagnosed with breast cancer: The Breast Cancer Health Disparities Study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3280. doi:10.1158/1538-7445.AM2017-3280

Effects of long-term ethanol consumption and Aldh1b1 depletion on intestinal tumourigenesis in mice.

Ethanol and its metabolite acetaldehyde have been classified as carcinogens for the upper aerodigestive tract, liver, breast, and colorectum. Whereas mechanisms related to oxidative stress and Cyp2e1 induction seem to prevail in the liver, and acetaldehyde has been proposed to play a crucial role in the upper aerodigestive tract, pathological mechanisms in the colorectum have not yet been clarified. Moreover, all evidence for a pro-carcinogenic role of ethanol in colorectal cancer is derived from correlations observed in epidemiological studies or from rodent studies with additional carcinogen application or tumour suppressor gene inactivation. In the current study, wild-type mice and mice with depletion of aldehyde dehydrogenase 1b1 (Aldh1b1), an enzyme which has been proposed to play an important role in acetaldehyde detoxification in the intestines, received ethanol in drinking water for 1 year. Long-term ethanol consumption led to intestinal tumour development in wild-type and Aldh1b1-depleted mice, but no intestinal tumours were observed in water-treated controls. Moreover, a significant increase in DNA damage was detected in the large intestinal epithelium of ethanol-treated mice of both genotypes compared with the respective water-treated groups, along with increased proliferation of the small and large intestinal epithelium. Aldh1b1 depletion led to increased plasma acetaldehyde levels in ethanol-treated mice, to a significant aggravation of ethanol-induced intestinal hyperproliferation, and to more advanced features of intestinal tumours, but it did not affect intestinal tumour incidence. These data indicate that ethanol consumption can initiate intestinal tumourigenesis without any additional carcinogen treatment or tumour suppressor gene inactivation, and we provide evidence for a role of Aldh1b1 in protection of the intestines from ethanol-induced damage, as well as for both carcinogenic and tumour-promoting functions of acetaldehyde, including increased progression of ethanol-induced tumours. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Altering substrate specificity of aldehyde dehydrogenase 3A1 to enhance acetaldehyde metabolism, in vivo (585.9)

Acetaldehyde is a well‐know cytotoxin, and carcinogen. Detoxification of acetaldehyde, which is mainly carried out by the mitochondrial aldehyde dehydrogenase 2 (ALDH2), is crucial to human health. High levels of acetaldehydes are present in the saliva and in circulation after ethanol ingestion and likely contribute to the high incidence of esophageal cancer in alcoholics and in carriers of a common ALDH2 mutation (ALDH2*2) found in 540 million East Asians. Acetaldehyde is also partly responsible for behavior impairment following ethanol consumption. Therefore, means to accelerate acetaldehyde clearance will be beneficial for human health. We previously identified an activator of ALDH2 that accelerated acetaldehyde metabolism. Here we describe the recruitment of an additional ALDH isozyme, ALDH3A1, to accelerate the clearance of acetaldehyde by the use of a small molecule. ALDH2 is the main isozyme that metabolizes acetaldehyde in liver. In contrast, ALDH3A1, which is present in the stomach and mucosa of the esophagus, normally metabolizes aromatic aldehydes and is ineffective in metabolizing acetaldehyde. We identified a small molecule activator, Alda‐89, which increased the Kcat of ALDH3A1 for acetaldehyde and enhanced its catalysis. We showed that when given together with an ALDH2‐specific activator, Alda‐1, Alda‐89 induced a rapid reduction in blood ethanol and acetaldehyde levels after acute ingestion of ethanol in mice. Importantly, treatment with Alda‐1 and Alda‐89 also reduced behavioral impairment in both wild‐type and ALDH2*2 mice. The ability to recruit an enzyme to metabolize a substrate that it usually does not metabolize using a pharmacological agent may represent a novel strategy to increase metabolism of toxic agents, in vivo.Grant Funding Source: Supported by NIH AA11147 to DM‐R

Acetaldehyde Detoxification Using Resting Cells of Recombinant Escherichia coli Overexpressing Acetaldehyde Dehydrogenase

Acetaldehyde dehydrogenase (E.C. 1.2.1.10) plays a key role in the acetaldehyde detoxification. The recombinant Escherichia coli cells producing acetaldehyde dehydrogenase (ist-ALDH) were applied as whole-cell biocatalysts for biodegradation of acetaldehyde. Response surface methodology (RSM) was employed to enhance the production of recombinant ist-ALDH. Under the optimum culture conditions containing 20.68 h post-induction time, 126.75 mL medium volume and 3 % (v/v) inoculum level, the maximum ist-ALDH activity reached 496.65 ± 0.81 U/mL, resulting in 12.5-fold increment after optimization. Furthermore, the optimum temperature and pH for the catalytic activity of wet cells were 40 °C and pH 9.5, respectively. The biocatalytic activity was improved 80 % by permeabilizing the recombinant cells with 0.075 % (v/v) Triton X-100. When using 2 mmol/L NAD(+) as coenzyme, the permeabilized cells could catalyze 98 % of acetaldehyde within 15 min. The results indicated that the recombinant E. coli with high productivity of ist-ALDH might be highly efficient and easy-to-make biocatalysts for acetaldehyde detoxification.

Genotoxic consequences of endogenous aldehydes on mouse haematopoietic stem cell function

Haematopoietic stem cells (HSCs) regenerate blood cells throughout the lifespan of an organism. With age, the functional quality of HSCs declines, partly owing to the accumulation of damaged DNA. However, the factors that damage DNA and the protective mechanisms that operate in these cells are poorly understood. We have recently shown that the Fanconi anaemia DNA-repair pathway counteracts the genotoxic effects of reactive aldehydes. Mice with combined inactivation of aldehyde catabolism (through Aldh2 knockout) and the Fanconi anaemia DNA-repair pathway (Fancd2 knockout) display developmental defects, a predisposition to leukaemia, and are susceptible to the toxic effects of ethanol-an exogenous source of acetaldehyde. Here we report that aged Aldh2(-/-) Fancd2(-/-) mutant mice that do not develop leukaemia spontaneously develop aplastic anaemia, with the concomitant accumulation of damaged DNA within the haematopoietic stem and progenitor cell (HSPC) pool. Unexpectedly, we find that only HSPCs, and not more mature blood precursors, require Aldh2 for protection against acetaldehyde toxicity. Additionally, the aldehyde-oxidizing activity of HSPCs, as measured by Aldefluor stain, is due to Aldh2 and correlates with this protection. Finally, there is more than a 600-fold reduction in the HSC pool of mice deficient in both Fanconi anaemia pathway-mediated DNA repair and acetaldehyde detoxification. Therefore, the emergence of bone marrow failure in Fanconi anaemia is probably due to aldehyde-mediated genotoxicity restricted to the HSPC pool. These findings identify a new link between endogenous reactive metabolites and DNA damage in HSCs, and define the protective mechanisms that counteract this threat.