bims-toxgon 2023-05-28 papers

bims-toxgon

Biomed News

on Toxoplasma gondii metabolism

Issue of 2023–05–28
twelve papers selected by
Lakesh Kumar, BITS Pilani

Histone variant H2B.Z acetylation is necessary for maintenance of Toxoplasma gondii biological fitness.
Toxoplasma gondii infection: novel emerging therapeutic targets.
Egress Regulatory Factors: How Toxoplasma Exits from Infected Cells?
Fostering Innovation to solve the biomechanics of Microbe-Host Interactions: Focus on the adhesive forces underlying Apicomplexa parasite biology.
Apicomplexan parasites are attenuated by low-energy electron irradiation in an automated microfluidic system and protect against infection with Toxoplasma gondii.
Histone deacetylase (HDACs) inhibitors: Clinical applications.
MAP4K3 inhibits Sirtuin-1 to repress the LKB1-AMPK pathway to promote amino acid-dependent activation of the mTORC1 complex.
Bacterial structural genomics target enabled by a recently discovered potent fungal acetyl-CoA synthetase inhibitor.
Post-translational modifications of histones: Mechanisms, biological functions, and therapeutic targets.
Pyruvate dehydrogenase complex integrates the metabolome and epigenome in CD8+ memory T cell differentiation in vitro.
AMPK is a mechano-metabolic sensor linking cell adhesion and mitochondrial dynamics to Myosin-dependent cell migration.
Redefining the role of AMPK in autophagy and the energy stress response.

Biochim Biophys Acta Gene Regul Mech. 2023 May 20. pii: S1874-9399(23)00038-X. [Epub ahead of print]1866(3): 194943

Histone variant H2B.Z acetylation is necessary for maintenance of Toxoplasma gondii biological fitness.

Laura Vanagas, Daniela Muñoz, Constanza Cristaldi, Agustina Ganuza, Rosario Nájera, Mabel C Bonardi, Valeria R Turowski, Fanny Guzman, Bin Deng, Kami Kim, William J Sullivan, Sergio O Angel.

  Through regulation of DNA packaging, histone proteins are fundamental to a wide array of biological processes. A variety of post-translational modifications (PTMs), including acetylation, constitute a proposed histone code that is interpreted by "reader" proteins to modulate chromatin structure. Canonical histones can be replaced with variant versions that add an additional layer of regulatory complexity. The protozoan parasite Toxoplasma gondii is unique among eukaryotes in possessing a novel variant of H2B designated H2B.Z. The combination of PTMs and the use of histone variants are important for gene regulation in T. gondii, offering new targets for drug development. In this work, T. gondii parasites were generated in which the 5 N-terminal acetylatable lysines in H2B.Z were mutated to either alanine (c-Myc-A) or arginine (c-Myc-R). The c-Myc-A mutant displayed no phenotype over than a mild defect in its ability to kill mice. The c-Myc-R mutant presented an impaired ability to grow and an increase in differentiation to latent bradyzoites. The c-Myc-R mutant was also more sensitive to DNA damage, displayed no virulence in mice, and provided protective immunity against future infection. While nucleosome composition was unaltered, key genes were abnormally expressed during in vitro bradyzoite differentiation. Our results show that regulation of the N-terminal positive charge patch of H2B.Z is important for these processes. We also show that acetylated N-terminal H2B.Z interacts with some unique proteins compared to its unacetylated counterpart; the acetylated peptide pulled down proteins associated with chromosome maintenance/segregation and cell cycle, suggesting a link between H2B.Z acetylation status and mitosis.

Keywords:  Cell cycle; Differentiation; Mitosis; Positive charge patch; Virulence

DOI:  https://doi.org/10.1016/j.bbagrm.2023.194943
Expert Opin Ther Targets. 2023 May 24. 1-12

Toxoplasma gondii infection: novel emerging therapeutic targets.

Joachim Müller, Andrew Hemphill.

   INTRODUCTION: Toxoplasmosis constitutes a challenge for public health, animal production, and welfare. So far, only a limited panel of drugs has been marketed for clinical applications. In addition to classical screening, the investigation of unique targets of the parasite may lead to the identification of novel drugs.
AREAS COVERED: Herein, the authors describe the methodology to identify novel drug targets in Toxoplasma gondii and review the literature with a focus on the last two decades.
EXPERT OPINION: Over the last two decades, the investigation of essential proteins of T. gondii as potential drug targets has fostered the hope of identifying novel compounds for the treatment of toxoplasmosis. Despite good efficacies in vitro, only a few classes of these compounds are effective in suitable rodent models, and none has cleared the hurdle to applications in humans. This shows that target-based drug discovery is in no way better than classical screening approaches. In both cases, off-target effects and adverse side effects in the hosts must be considered. Proteomics-driven analyses of parasite- and host-derived proteins that physically bind drug candidates may constitute a suitable tool to characterize drug targets, irrespectively of the drug discovery methods.

Keywords:  Apicomplexa; drug development; drug targets; host–parasite interactions; proteomics

DOI:  https://doi.org/10.1080/14728222.2023.2217353
Pathogens. 2023 May 04. pii: 679. [Epub ahead of print]12(5):

Egress Regulatory Factors: How Toxoplasma Exits from Infected Cells?

Yujie Diao, Yong Yao, Saeed El-Ashram, Maohong Bian.

  Toxoplasma gondii is an obligatory intracellular protozoan in the family Apicomplexa. It infects almost one-third of the world's population and causes toxoplasmosis, a prevalent disease. The parasite's egress from infected cells is a key step in the pathology caused by T. gondii. Moreover, T. gondii's continuous infection relies heavily on its capacity to migrate from one cell to another. Many pathways are involved in T. gondii egress. Individual routes may be modified to respond to various environmental stimuli, and many paths can converge. Regardless of the stimuli, the relevance of Ca2+ as a second messenger in transducing these signals, and the convergence of various signaling pathways in the control of motility and, ultimately, egress, is well recognized. This review attempts to outline intra- and extra-parasitic regulators that mediate T. gondii egress, and provides insight into potential clinical interventions and research.

Keywords:  Egress; Pathogenesis; Toxoplasma gondii

DOI:  https://doi.org/10.3390/pathogens12050679
Biol Cell. 2023 May 25.

Fostering Innovation to solve the biomechanics of Microbe-Host Interactions: Focus on the adhesive forces underlying Apicomplexa parasite biology.

Luis Vigetti, Isabelle Tardieux.

  The protozoa Toxoplasma gondii and Plasmodium spp., are preeminent members of the Apicomplexa parasitic phylum in large part due to their public health and economic impact. Hence, they serve as model unicellular eukaryotes with which to explore the repertoire of molecular and cellular strategies that specific developmental morphotypes deploy to timely adjust to their host(s) in order to perpetuate. In particular, host tissue- and cell-invasive morphotypes termed zoites alternate extracellular and intracellular lifestyles, thereby sensing and reacting to a wealth of host-derived biomechanical cues over their partnership. In the recent years, biophysical tools especially related to real time force measurement have been introduced, teaching us how creative are these microbes to shape a unique motility system that powers fast gliding through a variety of extracellular matrices, across cellular barriers, in vascular systems or into host cells. Equally performant was this toolkit to start illuminating how parasites manipulate their hosting cell adhesive and rheological properties to their advantage. In this review, besides highlighting major discoveries along the way, we discuss the most promising development, synergy and multimodal integration in active noninvasive force microscopy methods. These should in the near future unlock current limitations and allow capturing, from molecules to tissues, the many biomechanical and biophysical interplays over the dynamic host and microbe partnership. This article is protected by copyright. All rights reserved.

Keywords:  Apicomplexa; adhesive strength; force microscopy; live imaging; mechanobiology; parasitism; technical synergy

DOI:  https://doi.org/10.1111/boc.202300016
Parasitol Res. 2023 May 26.

Apicomplexan parasites are attenuated by low-energy electron irradiation in an automated microfluidic system and protect against infection with Toxoplasma gondii.

Julia Finkensieper, Florian Mayerle, Zaida Rentería-Solís, Jasmin Fertey, Gustavo R Makert, Franziska Lange, Joana Besecke, Simone Schopf, Andre Poremba, Ulla König, Bastian Standfest, Martin Thoma, Arwid Daugschies, Sebastian Ulbert.

  Radiation-attenuated intracellular parasites are promising immunization strategies. The irradiated parasites are able to invade host cells but fail to fully replicate, which allows for the generation of an efficient immune response. Available radiation technologies such as gamma rays require complex shielding constructions and are difficult to be integrated into pharmaceutical production processes. In this study, we evaluated for the first time low-energy electron irradiation (LEEI) as a method to generate replication-deficient Toxoplasma gondii and Cryptosporidium parvum. Similar to other radiation technologies, LEEI mainly damages nucleic acids; however, it is applicable in standard laboratories. By using a novel, continuous, and microfluidic-based LEEI process, tachyzoites of T. gondii and oocysts of C. parvum were irradiated and subsequently analyzed in vitro. The LEEI-treated parasites invaded host cells but were arrested in intracellular replication. Antibody-based analysis of surface proteins revealed no significant structural damage due to LEEI. Similarly, excystation rates of sporozoites from irradiated C. parvum oocysts were similar to those from untreated controls. Upon immunization of mice, LEEI-attenuated T. gondii tachyzoites induced high levels of antibodies and protected the animals from acute infection. These results suggest that LEEI is a useful technology for the generation of attenuated Apicomplexan parasites and has potential for the development of anti-parasitic vaccines.

Keywords:  Attenuation; Cryptosporidium; Irradiation; Toxoplasma; Vaccine

DOI:  https://doi.org/10.1007/s00436-023-07880-w
Prog Mol Biol Transl Sci. 2023 ;pii: S1877-1173(23)00056-X. [Epub ahead of print]198 119-152

Histone deacetylase (HDACs) inhibitors: Clinical applications.

Kumar D Shanmukha, Harikrishnareddy Paluvai, Santosh K Lomada, Mahesh Gokara, Suresh K Kalangi.

  Histone Deacetylases (HDACs) deacetylate lysine residues in histone and non-histone proteins. HDACs have been implicated in several diseases, including cancer, neurodegeneration, and cardiovascular disease. HDACs play an essential role in gene transcription, cell survival, growth, and proliferation, with histone hypoacetylation as one of the critical downstream signatures. HDAC inhibitors (HDACi) regulate gene expression epigenetically by restoring acetylation levels. Contrarily, only few HDACi have received FDA approval, and the majority are presently undergoing clinical trials to ascertain their effectiveness in the prevention and treatment of disease. In this book chapter, we give a detailed list of HDAC classes, and their functions in advancing diseases like cancer, cardiovascular, and neurodegeneration. Furthermore we touch upon novel and promising HDACi therapy approaches in the relevance of the current clinical scenario.

Keywords:  Cancer; Cardiovascular; HDACi; HDACs; Neurodegeneration

DOI:  https://doi.org/10.1016/bs.pmbts.2023.02.011
Life Sci Alliance. 2023 08;pii: e202201525. [Epub ahead of print]6(8):

MAP4K3 inhibits Sirtuin-1 to repress the LKB1-AMPK pathway to promote amino acid-dependent activation of the mTORC1 complex.

Mary Rose Branch, Cynthia L Hsu, Kohta Ohnishi, Wen-Chuan Shen, Elian Lee, Jill Meisenhelder, Brett Winborn, Bryce L Sopher, J Paul Taylor, Tony Hunter, Albert R La Spada.

mTORC1 is the key rheostat controlling the cellular metabolic state. Of the various inputs to mTORC1, the most potent effector of intracellular nutrient status is amino acid supply. Despite an established role for MAP4K3 in promoting mTORC1 activation in the presence of amino acids, the signaling pathway by which MAP4K3 controls mTORC1 activation remains unknown. Here, we examined the process of MAP4K3 regulation of mTORC1 and found that MAP4K3 represses the LKB1-AMPK pathway to achieve robust mTORC1 activation. When we sought the regulatory link between MAP4K3 and LKB1 inhibition, we discovered that MAP4K3 physically interacts with the master nutrient regulatory factor sirtuin-1 (SIRT1) and phosphorylates SIRT1 to repress LKB1 activation. Our results reveal the existence of a novel signaling pathway linking amino acid satiety with MAP4K3-dependent suppression of SIRT1 to inactivate the repressive LKB1-AMPK pathway and thereby potently activate the mTORC1 complex to dictate the metabolic disposition of the cell.

DOI: https://doi.org/10.26508/lsa.202201525
Acta Crystallogr F Struct Biol Commun. 2023 Jun 01.

Bacterial structural genomics target enabled by a recently discovered potent fungal acetyl-CoA synthetase inhibitor.

Nicholas D DeBouver, Madison J Bolejack, Taiwo E Esan, Damian J Krysan, Timothy J Hagen, Jan Abendroth.

  The compound ethyl-adenosyl monophosphate ester (ethyl-AMP) has been shown to effectively inhibit acetyl-CoA synthetase (ACS) enzymes and to facilitate the crystallization of fungal ACS enzymes in various contexts. In this study, the addition of ethyl-AMP to a bacterial ACS from Legionella pneumophila resulted in the determination of a co-crystal structure of this previously elusive structural genomics target. The dual functionality of ethyl-AMP in both inhibiting ACS enzymes and promoting crystallization establishes its significance as a valuable resource for advancing structural investigations of this class of proteins.

Keywords:  Legionella pneumophila; SSGCID; acetyl-coenzyme A synthetase; ligases; structural genomics

DOI:  https://doi.org/10.1107/S2053230X23003801
MedComm (2020). 2023 Jun;4(3): e292

Post-translational modifications of histones: Mechanisms, biological functions, and therapeutic targets.

Ruiqi Liu, Jiajun Wu, Haiwei Guo, Weiping Yao, Shuang Li, Yanwei Lu, Yongshi Jia, Xiaodong Liang, Jianming Tang, Haibo Zhang.

  Histones are DNA-binding basic proteins found in chromosomes. After the histone translation, its amino tail undergoes various modifications, such as methylation, acetylation, phosphorylation, ubiquitination, malonylation, propionylation, butyrylation, crotonylation, and lactylation, which together constitute the "histone code." The relationship between their combination and biological function can be used as an important epigenetic marker. Methylation and demethylation of the same histone residue, acetylation and deacetylation, phosphorylation and dephosphorylation, and even methylation and acetylation between different histone residues cooperate or antagonize with each other, forming a complex network. Histone-modifying enzymes, which cause numerous histone codes, have become a hot topic in the research on cancer therapeutic targets. Therefore, a thorough understanding of the role of histone post-translational modifications (PTMs) in cell life activities is very important for preventing and treating human diseases. In this review, several most thoroughly studied and newly discovered histone PTMs are introduced. Furthermore, we focus on the histone-modifying enzymes with carcinogenic potential, their abnormal modification sites in various tumors, and multiple essential molecular regulation mechanism. Finally, we summarize the missing areas of the current research and point out the direction of future research. We hope to provide a comprehensive understanding and promote further research in this field.

Keywords:  acetylation; cancer; methylation; phosphorylation; post‐translational modifications

DOI:  https://doi.org/10.1002/mco2.292
Res Sq. 2023 May 10. pii: rs.3.rs-2838359. [Epub ahead of print]

Pyruvate dehydrogenase complex integrates the metabolome and epigenome in CD8+ memory T cell differentiation in vitro.

Tatiana Tarasenko, Payal Banerjee, Julio Gomez-Rodriguez, Derek Gildea, Suiyuan Zhang, Tyra Wolfsberg, Lisa Jenkins, Nisc Comparative Sequencing Program, Peter McGuire.

Modulation of metabolic flux through pyruvate dehydrogenase complex (PDC) plays an important role in T cell activation and differentiation. PDC sits at the transition between glycolysis and the tricarboxylic acid cycle and is a major producer of acetyl-CoA, marking it as a potential metabolic and epigenetic node. To understand the role of pyruvate dehydrogenase complex in T cell differentiation, we generated mice deficient in T cell pyruvate dehydrogenase E1A ( Pdha ) subunit using a CD4-cre recombinase-based strategy. Herein, we show that genetic ablation of PDC activity in T cells ( TPdh -/- ) leads to marked perturbations in glycolysis, the tricarboxylic acid cycle, and OXPHOS. TPdh -/- T cells became dependent upon substrate level phosphorylation via glycolysis, secondary to depressed OXPHOS. Due to the block of PDC activity, histone acetylation was also reduced, including H3K27, a critical site for CD8 + T M differentiation. Transcriptional and functional profiling revealed abnormal CD8 + T M differentiation in vitro. Collectively, our data indicate that PDC integrates the metabolome and epigenome in CD8 + memory T cell differentiation. Targeting this metabolic and epigenetic node can have widespread ramifications on cellular function.

DOI: https://doi.org/10.21203/rs.3.rs-2838359/v1
Nat Commun. 2023 May 22. 14(1): 2740

AMPK is a mechano-metabolic sensor linking cell adhesion and mitochondrial dynamics to Myosin-dependent cell migration.

Eva Crosas-Molist, Vittoria Graziani, Oscar Maiques, Pahini Pandya, Joanne Monger, Remi Samain, Samantha L George, Saba Malik, Jerrine Salise, Valle Morales, Adrien Le Guennec, R Andrew Atkinson, Rosa M Marti, Xavier Matias-Guiu, Guillaume Charras, Maria R Conte, Alberto Elosegui-Artola, Mark Holt, Victoria Sanz-Moreno.

Cell migration is crucial for cancer dissemination. We find that AMP-activated protein kinase (AMPK) controls cell migration by acting as an adhesion sensing molecular hub. In 3-dimensional matrices, fast-migrating amoeboid cancer cells exert low adhesion/low traction linked to low ATP/AMP, leading to AMPK activation. In turn, AMPK plays a dual role controlling mitochondrial dynamics and cytoskeletal remodelling. High AMPK activity in low adhering migratory cells, induces mitochondrial fission, resulting in lower oxidative phosphorylation and lower mitochondrial ATP. Concurrently, AMPK inactivates Myosin Phosphatase, increasing Myosin II-dependent amoeboid migration. Reducing adhesion or mitochondrial fusion or activating AMPK induces efficient rounded-amoeboid migration. AMPK inhibition suppresses metastatic potential of amoeboid cancer cells in vivo, while a mitochondrial/AMPK-driven switch is observed in regions of human tumours where amoeboid cells are disseminating. We unveil how mitochondrial dynamics control cell migration and suggest that AMPK is a mechano-metabolic sensor linking energetics and the cytoskeleton.

DOI: https://doi.org/10.1038/s41467-023-38292-0
Nat Commun. 2023 May 24. 14(1): 2994

Redefining the role of AMPK in autophagy and the energy stress response.

Ji-Man Park, Da-Hye Lee, Do-Hyung Kim.

Autophagy maintains cellular homeostasis during low energy states. According to the current understanding, glucose-depleted cells induce autophagy through AMPK, the primary energy-sensing kinase, to acquire energy for survival. However, contrary to the prevailing concept, our study demonstrates that AMPK inhibits ULK1, the kinase responsible for autophagy initiation, thereby suppressing autophagy. We found that glucose starvation suppresses amino acid starvation-induced stimulation of ULK1-Atg14-Vps34 signaling via AMPK activation. During an energy crisis caused by mitochondrial dysfunction, the LKB1-AMPK axis inhibits ULK1 activation and autophagy induction, even under amino acid starvation. Despite its inhibitory effect, AMPK protects the ULK1-associated autophagy machinery from caspase-mediated degradation during energy deficiency, preserving the cellular ability to initiate autophagy and restore homeostasis once the stress subsides. Our findings reveal that dual functions of AMPK, restraining abrupt induction of autophagy upon energy shortage while preserving essential autophagy components, are crucial to maintain cellular homeostasis and survival during energy stress.

DOI: https://doi.org/10.1038/s41467-023-38401-z