Alpha-Gal Inactivated Heart Valve Bioprostheses Exhibit an Anti-Calcification Propensity Similar to Knockout Tissues<sup />

Naso, Filippo; Stefanelli, U; Buratto, Edward; Lazzari, Giovanna; Perota, Andrea; Galli, Cesare; Gandaglia, Alessandro

doi:10.1089/ten.tea.2016.0474

Cited by 33 publications

(37 citation statements)

References 30 publications

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“…Serum was stored at −80°C until further use for purification 17 . Double knockout (DKO) pigs were obtained by targeting the swine CMAH gene using TALEN 28 on the GGTA12/2 background (obtained from Dr Soulillou through Dr D. H. Sachs, Massachusetts General Hospital, Boston, MA, USA) 29 and by somatic cell nuclear transfer 28,30 . The CKO pigs were generated in Dr C. Galli's laboratory 31 by breeding DKO male founders to WT sows.…”

Section: Methodsmentioning

confidence: 99%

Liquid chromatography–tandem mass spectrometry glycoproteomic study of porcine IgG and detection of subtypes

Battellino

Bacala

Gigolyk

et al. 2021

Rapid Comm Mass Spectrometry

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Rationale While high‐throughput proteomic methods have been widely applied to monoclonal antibodies and human immunoglobulin gamma (IgG) samples, less information is available on porcine IgG. As pigs are considered one of the most suitable species for xenotransplantation, it is important to characterize IgG amino acid sequences and glycosylation profiles, which is the focus of this study. Methods Three different purified porcine IgG samples, including wild‐type and knockout species, were digested with trypsin and enriched for glycopeptides. Digestion mixtures were spiked with a mixture of six standard peptides. Analysis was performed using electrospray ionization liquid chromatography–tandem mass spectrometry (MS/MS) in standard MS/MS data‐dependent acquisition mode on a hybrid triple quadrupole time‐of‐flight mass spectrometer. Results To facilitate the classification of subtypes detected experimentally, UniprotKB database entries were organized using comparative alignment scores. Sequences were grouped based on 11 different subtypes as translated from GenBank entries. Proteomic searches were accomplished automatically using specialized software, whereas glycoprotein searches were performed manually by monitoring the extracted chromatograms of diagnostic MS/MS glycan fragments and studying their corresponding mass spectra; 40–50 non‐glycosylated peptides and 4–5 glycosylated peptides were detected in each sample, with several glycoforms per sequence. Conclusions Proteomic analysis of porcine IgG is complicated by factors such as the presence of several subtypes, redundant heavy chain (HC) sequences in protein databases, and the lack of consistent cross‐referencing between databases. Aligning and comparing HC sequences were necessary to eliminate redundancy. This study highlights the complexity of pig IgG and shows the importance of MS in proteomics and glycoproteomics.

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Section: Methodsmentioning

confidence: 99%

Liquid chromatography–tandem mass spectrometry glycoproteomic study of porcine IgG and detection of subtypes

Battellino

Bacala

Gigolyk

et al. 2021

Rapid Comm Mass Spectrometry

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“…There is growing interest in using Gal-free animal tissues from Gal-knockout pigs (GTKO) to reduce anti-Gal antibody-associated tissue calcification [18, 21, 22, 37, 38]. While the potential advantage of Gal-free tissue has not been clinically demonstrated, our previous studies have shown that glutaraldehyde-fixed GTKO and standard porcine pericardial tissues have same general morphology, collagen content, and mechanical strength [23].…”

Section: Discussionmentioning

confidence: 99%

A Durable Porcine Pericardial Surgical Bioprosthetic Heart Valve: a Proof of Concept

Rahmani

McGregor

Byrne

et al. 2019

J. of Cardiovasc. Trans. Res.

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Bioprosthetic leaflets made from animal tissues are used in the majority of surgical and transcatheter cardiac valve replacements. This study develops a new surgical bioprosthesis, using porcine pericardial leaflets. Porcine pericardium was obtained from genetically engineered pigs with a mutation in the GGTA-1 gene (GTKO) and fixed in 0.6% glutaraldehyde, and used to develop a new surgical valve design. The valves underwent in vitro hydrodynamic test in a pulse duplicator and high-cycled accelerated wear testing and were evaluated for acute haemodynamics and thrombogenicity in a juvenile sheep implant study for 48 h. The porcine surgical pericardial heart valves (pSPHVs) exhibited excellent hydrodynamics and reached 200 million cycles of in vitro durability, with no observable damage. Juvenile sheep implants demonstrated normal valve function with no acute thrombogenic response for either material. The pSPHV incorporates a minimalistic construction method using a tissue-to-tissue design to cover the stent. This new design is a proof of concept alternative to the use of bovine pericardium and synthetic fabric in surgical bioprosthetic heart valves.

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“…8 The resulting humoral response leads to activation of the complement system, triggering endothelial cell dysfunction, platelet aggregation, and promotes calcification. 2,[9][10][11] Although the presented study of Veraar and colleagues 5 did not explore any relationship between the degree of immunogenic response and valve durability, other groups were able to show an association of anti-alpha-gal antibodies and premature bioprosthetic valve degeneration. 12 Furthermore, several experimental studies demonstrated a connection between anti-alpha-gal antibodies and the calcification process in valvular bioprosthesis.…”

mentioning

confidence: 88%

Commentary: From Old World monkeys to New World humans—Evolved protection from tick bites and bioprosthetic material

2021

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Alpha-Gal Inactivated Heart Valve Bioprostheses Exhibit an Anti-Calcification Propensity Similar to Knockout Tissues

Cited by 33 publications

References 30 publications

Liquid chromatography–tandem mass spectrometry glycoproteomic study of porcine IgG and detection of subtypes

Liquid chromatography–tandem mass spectrometry glycoproteomic study of porcine IgG and detection of subtypes

A Durable Porcine Pericardial Surgical Bioprosthetic Heart Valve: a Proof of Concept

Commentary: From Old World monkeys to New World humans—Evolved protection from tick bites and bioprosthetic material

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