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Molecular Cancer Feb 2022Dendritic cells (DCs) are central for the initiation and regulation of innate and adaptive immunity in the tumor microenvironment. As such, many kinds of DC-targeted...
BACKGROUND
Dendritic cells (DCs) are central for the initiation and regulation of innate and adaptive immunity in the tumor microenvironment. As such, many kinds of DC-targeted vaccines have been developed to improve cancer immunotherapy in numerous clinical trials. Targeted delivery of antigens and adjuvants to DCs in vivo represents an important approach for the development of DC vaccines. However, nonspecific activation of systemic DCs and the preparation of optimal immunodominant tumor antigens still represent major challenges.
METHODS
We loaded the immunogenic cell death (ICD) inducers human neutrophil elastase (ELANE) and Hiltonol (TLR3 agonist) into α-lactalbumin (α-LA)-engineered breast cancer-derived exosomes to form an in situ DC vaccine (HELA-Exos). HELA-Exos were identified by transmission electron microscopy, nanoscale flow cytometry, and Western blot analysis. The targeting, killing, and immune activation effects of HELA-Exos were evaluated in vitro. The tumor suppressor and immune-activating effects of HELA-Exos were explored in immunocompetent mice and patient-derived organoids.
RESULTS
HELA-Exos possessed a profound ability to specifically induce ICD in breast cancer cells. Adequate exposure to tumor antigens and Hiltonol following HELA-Exo-induced ICD of cancer cells activated type one conventional DCs (cDC1s) in situ and cross-primed tumor-reactive CD8 T cell responses, leading to potent tumor inhibition in a poorly immunogenic triple negative breast cancer (TNBC) mouse xenograft model and patient-derived tumor organoids.
CONCLUSIONS
HELA-Exos exhibit potent antitumor activity in both a mouse model and human breast cancer organoids by promoting the activation of cDC1s in situ and thus improving the subsequent tumor-reactive CD8 T cell responses. The strategy proposed here is promising for generating an in situ DC-primed vaccine and can be extended to various types of cancers. Scheme 1. Schematic illustration of HELA-Exos as an in situ DC-primed vaccine for breast cancer. (A) Allogenic breast cancer-derived exosomes isolated from MDA-MB-231 cells were genetically engineered to overexpress α-LA and simultaneously loaded with the ICD inducers ELANE and Hiltonol (TLR3 agonist) to generate HELA-Exos. (B) Mechanism by which HELA-Exos activate DCs in situ in a mouse xenograft model ofTNBC. HELA-Exos specifically homed to the TME and induced ICD in cancer cells, which resulted in the increased release of tumor antigens, Hiltonol, and DAMPs, as well as the uptake of dying tumor cells by cDC1s. The activated cDC1s then cross-primed tumor-reactive CD8+ T cell responses. (C) HELA-Exos activated DCs in situ in the breast cancer patient PBMC-autologous tumor organoid coculture system.
ABBREVIATIONS
DCs: dendritic cells; α-LA: α-lactalbumin; HELA-Exos: Hiltonol-ELANE-α-LA-engineered exosomes; ICD: immunogenic cell death; ELANE: human neutrophil elastase; TLR3: Toll-like receptor 3; TNBC: triple-negative breast cancer; TME: tumor microenvironment; DAMPs: damage-associated molecular patterns; cDC1s: type 1 conventional dendritic cells; PBMCs: peripheral blood mononuclear cells.
Topics: Animals; Breast Neoplasms; Cancer Vaccines; Cell Line, Tumor; Dendritic Cells; Exosomes; Female; Humans; Leukocytes, Mononuclear; Mice; Tumor Microenvironment; Vaccines
PubMed: 35148751
DOI: 10.1186/s12943-022-01515-x -
Trends in Endocrinology and Metabolism:... Jun 2020This review details the physiologic roles of two insulin sensitizers, myo-inositol (MI) and d-chiro-inositol (DCI). In the human ovary, MI is a second messenger of... (Review)
Review
This review details the physiologic roles of two insulin sensitizers, myo-inositol (MI) and d-chiro-inositol (DCI). In the human ovary, MI is a second messenger of follicle-stimulating hormone (FSH) and DCI is an aromatase inhibitor. These activities allow a treatment for polycystic ovary syndrome (PCOS) to be defined based on the combined administration of MI and DCI, where the best MI:DCI ratio is 40:1. Moreover, MI enhances the effect of metformin and clomiphene on the fertility of PCOS women seeking pregnancy. As impaired intestinal transport may lead to unsuccessful inositol treatment, we also discuss new data on the use of alpha-lactalbumin to boost inositol absorption. Overall, the physiological activities of MI and DCI dictate the dosages and timing of inositol supplementation in the treatment of PCOS.
Topics: Animals; Female; Humans; Inositol; Polycystic Ovary Syndrome; Vitamin B Complex
PubMed: 32396844
DOI: 10.1016/j.tem.2020.02.002 -
Biomolecules Aug 2020α-Lactalbumin (α-LA) is a small (Mr 14,200), acidic (pI 4-5), Ca-binding protein. α-LA is a regulatory component of lactose synthase enzyme system functioning in the... (Review)
Review
α-Lactalbumin (α-LA) is a small (Mr 14,200), acidic (pI 4-5), Ca-binding protein. α-LA is a regulatory component of lactose synthase enzyme system functioning in the lactating mammary gland. The protein possesses a single strong Ca-binding site, which can also bind Mg, Mn, Na, K, and some other metal cations. It contains several distinct Zn-binding sites. Physical properties of α-LA strongly depend on the occupation of its metal binding sites by metal ions. In the absence of bound metal ions, α-LA is in the molten globule-like state. The binding of metal ions, and especially of Ca, increases stability of α-LA against the action of heat, various denaturing agents and proteases, while the binding of Zn to the Ca-loaded protein decreases its stability and causes its aggregation. At pH 2, the protein is in the classical molten globule state. α-LA can associate with membranes at neutral or slightly acidic pH at physiological temperatures. Depending on external conditions, α-LA can form amyloid fibrils, amorphous aggregates, nanoparticles, and nanotubes. Some of these aggregated states of α-LA can be used in practical applications such as drug delivery to tissues and organs. α-LA and some of its fragments possess bactericidal and antiviral activities. Complexes of partially unfolded α-LA with oleic acid are cytotoxic to various tumor and bacterial cells. α-LA in the cytotoxic complexes plays a role of a delivery carrier of cytotoxic fatty acid molecules into tumor and bacterial cells across the cell membrane. Perhaps in the future the complexes of α-LA with oleic acid will be used for development of new anti-cancer drugs.
Topics: Animals; Antineoplastic Agents; Humans; Hydrogen-Ion Concentration; Lactalbumin; Neoplasms; Oleic Acid
PubMed: 32825311
DOI: 10.3390/biom10091210 -
Nutrients Aug 2020Disturbed sleep may negatively influence physical health, cognitive performance, metabolism, and general wellbeing. Nutritional interventions represent a potential... (Randomized Controlled Trial)
Randomized Controlled Trial
BACKGROUND
Disturbed sleep may negatively influence physical health, cognitive performance, metabolism, and general wellbeing. Nutritional interventions represent a potential non-pharmacological means to increase sleep quality and quantity.
OBJECTIVE
(1) Identify an optimal suite of nutritional ingredients and (2) validate the effects of this suite utilising polysomnography, and cognitive and balance tests.
METHODS
The optimal and least optimal combinations of six ingredients were identified utilising 55 male participants and a Box-Behnken predictive model. To validate the model, 18 healthy, male, normal sleepers underwent three trials in a randomised, counterbalanced design: (1) optimal drink, (2) least optimal drink, or (3) placebo were provided before bed in a double-blinded manner. Polysomnography was utilised to measure sleep architecture. Cognitive performance, postural sway, and subjective sleep quality, were assessed 30 min after waking.
RESULTS
The optimal drink resulted in a significantly shorter sleep onset latency (9.9 ± 12.3 min) when compared to both the least optimal drink (26.1 ± 37.4 min) and the placebo drink (19.6 ± 32.0 min). No other measures of sleep, cognitive performance, postural sway, and subjective sleep quality were different between trials.
CONCLUSION
A combination of ingredients, optimised to enhance sleep, significantly reduced sleep onset latency. No detrimental effects on sleep architecture, subjective sleep quality or next day performance were observed.
Topics: Adenosine Monophosphate; Adult; Dietary Supplements; Double-Blind Method; Fruit and Vegetable Juices; Glutamates; Humans; Lactalbumin; Male; Polysomnography; Prunus avium; Sleep; Tryptophan; Valerian
PubMed: 32854375
DOI: 10.3390/nu12092579 -
Pharmacology, Biochemistry, and Behavior Jul 2024Milk varieties and specific proteins exhibit anxiolytic-like actions in mice and rats exposed to several tests, the most prominent being the elevated plus-maze.... (Review)
Review
Milk varieties and specific proteins exhibit anxiolytic-like actions in mice and rats exposed to several tests, the most prominent being the elevated plus-maze. Administrations of α-casein, its 91-100 (α-casozepine), 91-97, 91-93, and 91-92 fragments, the 60-69 fragment of β-casein, lactoferrin, β-lactotensin, wheylin-1, wheylin-2, and α-lactalbumin have been reported to increase open arm exploration relative to enclosed arm exploration. Anxiolytic-like actions have also been described for 91-93 and 91-92 fragments of α-casein, wheylin-1, α-lactalbumin, and lactoferrin in the open-field. Some effects appear to be mediated by the GABA receptor complex, since antagonists mitigated the anxiolytic-like actions of α-casein, the 91-92 fragment of α-casein, and wheylin-1. Other neurotransmitters purported to affect such behaviors include 5HT, dopamine, and neurotensin. Further research is needed to identify their neuropharmacological actions.
Topics: Animals; Anti-Anxiety Agents; Mice; Milk Proteins; Anxiety; Rats; Behavior, Animal; Humans; Caseins
PubMed: 38735399
DOI: 10.1016/j.pbb.2024.173789 -
Journal of Sleep Research Jan 2024Insufficient sleep is a growing global problem, with poor sleep associated with many negative health and performance outcomes. Previous reviews investigating the effect... (Review)
Review
Insufficient sleep is a growing global problem, with poor sleep associated with many negative health and performance outcomes. Previous reviews investigating the effect of diet on sleep have highlighted the amino acid tryptophan as a promising sleep-promoting nutrient, with the richest food source of tryptophan, ⍺-lactalbumin, requiring further investigation. Therefore, this systematic review aimed to review the existing evidence of association between ⍺-lactalbumin and sleep. Four electronic databases (CINAHL Complete, Embase, MEDLINE Complete, and SPORTDiscus with Full Text) were searched from database inception to March 2023, with primary research articles included if they contained α-lactalbumin as an independent variable, an outcome measure of sleep or sleepiness, and participants were ≥ 18 years old. Eight studies were reviewed, with four studies recruiting athletic populations (50%) and four recruiting healthy participants (50%). Sleep or sleepiness was measured objectively in six studies (75%), with two studies employing polysomnography and four utilizing actigraphy to assess sleep. Across the studies, 20-60 g of ⍺-lactalbumin was supplemented, with five studies (63%) observing a positive association between α-lactalbumin and sleep. Sleep-onset latency was the primary sleep metric improved following evening supplementation of α-lactalbumin (≤ 3.5 hr pre-sleep), with no studies observing any negative associations with sleep. Data from this review suggest that individuals that have difficulty initiating sleep may benefit most from pre-sleep α-lactalbumin supplementation. Further research is required to establish the effect that α-lactalbumin has on sleep architecture, through the use of more comprehensive sleep analysis tools such as portable electroencephalography or polysomnography, in combination with stringent dietary controls.
PubMed: 38185736
DOI: 10.1111/jsr.14141 -
Journal of the Science of Food and... Jun 2023Ultrasound-assisted glycation is a promising method for decreasing the allergenicity of α-lactalbumin (ALA). However, there is a lack of in vivo studies on the...
BACKGROUND
Ultrasound-assisted glycation is a promising method for decreasing the allergenicity of α-lactalbumin (ALA). However, there is a lack of in vivo studies on the allergenicity of ultrasound-assisted glycated ALA. In this study, the effects of the ultrasound-assisted glycation of ALA on the allergenicity and intestinal microflora were characterized using a BALB/c mouse model.
RESULTS
Increased immunoglobulin -G/ immunoglobulin-E (IgG/IgE) and interleukin-4/6 (IL-4/6) secretions, and reduced interferon-γ (IFN-γ) secretions were found in the serum of ALA sensitized and challenged, mice in comparison with a control group. However, there was no significant difference between the mice fed with ultrasound-assisted glycated ALA and the control group. Mice that were sensitized and challenged with ALA showed disrupted intestinal microflora, manifesting in significantly decreased Firmicutes and significantly increased Proteobacteria. It was found that 100ALA-gal could maintain the intestinal microflora of mice in a normal state. Pearson's rank correlation showed that Proteobacteria and Spirochaetota were correlated positively with the IL-4/IL-6 level and were correlated negatively with the expression of IFN-γ. Proteobacteria were also significantly positively correlated with IL-6 and negatively correlated with IFN-γ (P < 0.05).
CONCLUSION
These results suggested that ultrasound-assisted glycation on ALA can maintain the intestinal microflora in a normal state thus balancing the proportion of Th1/Th2 to decrease allergic reaction. © 2022 Society of Chemical Industry.
Topics: Animals; Mice; Allergens; Lactalbumin; Maillard Reaction; Interleukin-4; Interleukin-6; Immunoglobulin E; Interferon-gamma; Mice, Inbred BALB C
PubMed: 36303537
DOI: 10.1002/jsfa.12293 -
ACS Omega Nov 2022Alpha-lactalbumin (α-LA) and binding of zinc cations to protein were studied. Molecular characteristics of protein was determined by MALDI-TOF/MS and electrophoresis...
Alpha-lactalbumin (α-LA) and binding of zinc cations to protein were studied. Molecular characteristics of protein was determined by MALDI-TOF/MS and electrophoresis SDS-PAGE, and also, for complexes, it was determined by spectroscopic techniques (ATR-FT-IR and Raman) and microscopic techniques (SEM along with an EDX detector and also TEM). The pH dependence of zeta potential of α-LA was determined in saline solution. The zinc binding to the protein mechanism was investigated; zinc binding to protein kinetics, the molecular modeling by the DFT method, and electron microscopy (SEM and TEM) for microstructure observation were performed. The experiments performed indicate a quick binding process (equilibrium takes place after 2 min of incubation) which occurs onto the surface of α-LA. Zinc cations change the conformation of the protein and create spherical particles from the morphological point of view. DFT studies indicate the participation of acidic functional groups of the protein (aspartic acid and glutamic acid residues), and these have a decisive influence on the interaction with zinc cations. Application studies of general toxicity and cytotoxicity and bioavailability were conducted.
PubMed: 36340177
DOI: 10.1021/acsomega.2c03674 -
Journal of Dairy Science Mar 2021α-Lactalbumin (α-LA) and β-lactoglobulin (β-LG) were isolated from yak milk and identified by mass spectrometry. The variant of α-LA (L8IIC8) in yak milk had 123...
α-Lactalbumin (α-LA) and β-lactoglobulin (β-LG) were isolated from yak milk and identified by mass spectrometry. The variant of α-LA (L8IIC8) in yak milk had 123 amino acids, and the sequence differed from α-LA from bovine milk. The amino acid at site 71 was Asn (N) in domestic yak milk, but Asp (D) in bovine and wild yak milk sequences. Yak β-LG had 2 variants, β-LG A (P02754) and β-LG E (L8J1Z0). Both domestic yak and wild yak milk contained β-LG E, but it was absent in bovine milk. The amino acid at site 158 of β-Lg E was Gly (G) in yak but Glu (E) in bovine. The yak α-LA and β-LG secondary structures were slightly different from those in bovine milk. The denaturation temperatures of yak α-LA and β-LG were 52.1°C and 80.9°C, respectively. This study provides insights relevant to food functionality, food safety control, and the biological properties of yak milk products.
Topics: Animals; Cattle; Lactalbumin; Lactoglobulins; Milk; Milk Proteins; Whey Proteins
PubMed: 33358811
DOI: 10.3168/jds.2020-18546 -
International Journal of Molecular... Apr 2023Asthma is a heterogeneous inflammatory disease characterized by abnormalities in immune response. Due to the inherent complexity of the disease and the presence of... (Review)
Review
Asthma is a heterogeneous inflammatory disease characterized by abnormalities in immune response. Due to the inherent complexity of the disease and the presence of comorbidities, asthma control is often difficult to obtain. In asthmatic patients, an increased prevalence of irregular menstrual cycles, infertility, obesity, and insulin resistance has been reported. Given that these conditions are also common in patients with polycystic ovary syndrome (PCOS), we propose the definition of "asthma-PCOS overlap syndrome" to indicate a medical condition which shares characteristics of both diseases. The aim of this review is to analyze the links between asthma and PCOS and evaluate the therapeutic role of myo-inositol, a natural compound currently utilized in patients with PCOS, in the management of asthma patients.
Topics: Female; Humans; Polycystic Ovary Syndrome; Inositol; Insulin Resistance; Asthma
PubMed: 37108123
DOI: 10.3390/ijms24086959