bims-mricoa 2022-01-30 papers

Issue of 2022–01–30
four papers selected by
Merve Yavuz, Bilkent University

mSystems. 2022 Jan 25. e0103721

Global Analysis of Biomineralization Genes in Magnetospirillum magneticum AMB-1.

Hayley C McCausland, Kelly M Wetmore, Adam P Arkin, Arash Komeili.

Magnetotactic bacteria (MTB) are a phylogenetically diverse group of bacteria remarkable for their ability to biomineralize magnetite (Fe3O4) or greigite (Fe3S4) in organelles called magnetosomes. The majority of genes required for magnetosome formation are encoded by a magnetosome gene island (MAI). Most previous genetic studies of MTB have focused on the MAI, using screens to identify key MAI genes or targeted genetics to isolate specific genes and their function in one specific growth condition. This is the first study that has taken an unbiased approach to look at many different growth conditions to reveal key genes both inside and outside the MAI. Here, we conducted random barcoded transposon mutagenesis (RB-TnSeq) in Magnetospirillum magneticum AMB-1. We generated a library of 184,710 unique strains in a wild-type background, generating ∼34 mutant strains for each gene. RB-TnSeq also allowed us to determine the essential gene set of AMB-1 under standard laboratory growth conditions. To pinpoint novel genes that are important for magnetosome formation, we subjected the library to magnetic selection screens under varied growth conditions. We compared biomineralization under standard growth conditions to biomineralization under high-iron and anaerobic conditions, respectively. Strains with transposon insertions in the MAI gene mamT had an exacerbated biomineralization defect under both high-iron and anaerobic conditions compared to standard conditions, adding to our knowledge of the role of MamT in magnetosome formation. Mutants in an ex-MAI gene, amb4151, are more magnetic than wild-type cells under anaerobic conditions. All three of these phenotypes were validated by creating a markerless deletion strain of the gene and evaluating with TEM imaging. Overall, our results indicate that growth conditions affect which genes are required for biomineralization and that some MAI genes may have more nuanced functions than was previously understood. IMPORTANCE Magnetotactic bacteria (MTB) are a group of bacteria that can form nano-sized crystals of magnetic minerals. MTB are likely an important part of their ecosystems, because they can account for up to a third of the microbial biomass in an aquatic habitat and consume large amounts of iron, potentially impacting the iron cycle. The ecology of MTB is relatively understudied; however, the cell biology and genetics of MTB have been studied for decades. Here, we leverage genetic studies of MTB to inform environmental studies. We expand the genetic toolset for studying MTB in the lab and identify novel genes, or functions of genes, that have an impact on biomineralization.

Keywords: RB-TnSeq; biomineralization; magnetotactic bacteria

DOI: https://doi.org/10.1128/msystems.01037-21

Small. 2022 Jan 25. e2104763

Nanobugs as Drugs: Bacterial Derived Nanomagnets Enhance Tumor Targeting and Oncolytic Activity of HSV-1 Virus.

Faith H N Howard, Haider Al-Janabi, Priya Patel, Katie Cox, Emily Smith, Jayakumar Vadakekolathu, A Graham Pockley, Joe Conner, James F Nohl, Dan A Allwood, Cristal Collado-Rojas, Aneurin Kennerley, Sarah Staniland, Munitta Muthana.

The survival strategies of infectious organisms have inspired many therapeutics over the years. Indeed the advent of oncolytic viruses (OVs) exploits the uncontrolled replication of cancer cells for production of their progeny resulting in a cancer-targeting treatment that leaves healthy cells unharmed. Their success against inaccessible tumors however, is highly variable due to inadequate tumor targeting following systemic administration. Coassembling herpes simplex virus (HSV1716) with biocompatible magnetic nanoparticles derived from magnetotactic bacteria enables tumor targeting from circulation with magnetic guidance, protects the virus against neutralizing antibodies and thereby enhances viral replication within tumors. This approach additionally enhances the intratumoral recruitment of activated immune cells, promotes antitumor immunity and immune cell death, thereby inducing tumor shrinkage and increasing survival in a syngeneic mouse model of breast cancer by 50%. Exploiting the properties of such a nanocarrier, rather than tropism of the virus, for active tumor targeting offers an exciting, novel approach for enhancing the bioavailability and treatment efficacy of tumor immunotherapies for disseminated neoplasms.

Keywords: breast cancer; magnetic targeting; magnetosomes; nanomedicine; nanoparticles; oncolytic virotherapy

DOI: https://doi.org/10.1002/smll.202104763

Bioelectromagnetics. 2022 Feb;43(2): 119-143

Chain-Like Structures of Biogenic and Nonbiogenic Magnetic Nanoparticles in Vascular Tissues.

Svitlana Gorobets, Oksana Gorobets, Yuri Gorobets, Maryna Bulaievska.

In this paper, slices of organs from various organisms (animals, plants, fungi) were investigated by using atomic force microscopy and magnetic force microscopy to identify common features of localization of both biogenic and nonbiogenic magnetic nanoparticles. It was revealed that both biogenic and nonbiogenic magnetic nanoparticles are localized in the form of chains of separate nanoparticles or chains of conglomerates of nanoparticles in the walls of the capillaries of animals and the walls of the conducting tissue of plants and fungi. Both biogenic and nonbiogenic magnetic nanoparticles are embedded as a part of the transport system in multicellular organisms. In connection with this, a new idea of the function of biogenic magnetic nanoparticles is discussed, that the chains of biogenic magnetic nanoparticles and chains of conglomerates of biogenic magnetic nanoparticles represent ferrimagnetic organelles of a specific purpose. Besides, magnetic dipole-dipole interaction of biogenic magnetic nanoparticles with magnetically labeled drugs or contrast agents for magnetic resonance imaging should be considered when designing the drug delivery and other medical systems because biogenic magnetic nanoparticles in capillary walls will serve as the trapping centers for the artificial magnetic nanoparticles. The aggregates of both artificial and biogenic magnetic nanoparticles can be formed, contributing to the risk of vascular occlusion. Bioelectromagnetics. 43:119-143, 2022. © 2021 Bioelectromagnetics Society.

Keywords: biogenic magnetic nanoparticles; bioinformatics analysis; capillaries; ferrimagnetic organelles; magnetic force microscopy

DOI: https://doi.org/10.1002/bem.22390

PLoS One. 2022 ;17(1): e0262950

Biocompatible micro tweezers for 3D hydrogel organoid array mechanical characterization.

Soliman Alhudaithy, Kazunori Hoshino.

This study presents novel biocompatible Polydimethylsiloxane (PDMS)-based micromechanical tweezers (μTweezers) capable of the stiffness characterization and manipulation of hydrogel-based organoids. The system showed great potential for complementing established mechanical characterization methods such as Atomic Force Microscopy (AFM), parallel plate compression (PPC), and nanoindentation, while significantly reducing the volume of valuable hydrogels used for testing. We achieved a volume reduction of ~0.22 μl/sample using the μTweezers vs. ~157 μl/sample using the PPC, while targeting high-throughput measurement of widely adopted micro-mesoscale (a few hundred μm-1500 μm) 3D cell cultures. The μTweezers applied and measured nano-millinewton forces through cantilever' deflection with high linearity and tunability for different applications; the assembly is compatible with typical inverted optical microscopes and fit on standard tissue culture Petri dishes, allowing mechanical compression characterization of arrayed 3D hydrogel-based organoids in a high throughput manner. The average achievable output per group was 40 tests per hour, where 20 organoids and 20 reference images in one 35 mm petri dish were tested, illustrating efficient productivity to match the increasing demand on 3D organoids' applications. The changes in stiffness of collagen I hydrogel organoids in four conditions were measured, with ovarian cancer cells (SKOV3) or without (control). The Young's modulus of the control group (Control-day 0, E = 407± 146, n = 4) measured by PPC was used as a reference modulus, where the relative elastic compressive modulus of the other groups based on the stiffness measurements was also calculated (control-day 0, E = 407 Pa), (SKOV3-day 0, E = 318 Pa), (control-day 5, E = 528 Pa), and (SKOV3-day 5, E = 376 Pa). The SKOV3-embedded hydrogel-based organoids had more shrinkage and lowered moduli on day 0 and day 5 than controls, consistently, while SKOV3 embedded organoids increased in stiffness in a similar trend to the collagen I control from day 0 to day 5. The proposed method can contribute to the biomedical, biochemical, and regenerative engineering fields, where bulk mechanical characterization is of interest. The μTweezers will also provide attractive design and application concepts to soft membrane-micro 3D robotics, sensors, and actuators.

DOI: https://doi.org/10.1371/journal.pone.0262950