Combined Cononsolvency and Temperature Effects on Adsorbed PNIPAM Microgels.

The present study addresses the multiresponsive behavior of poly(N-isopropylacrylamide) (PNIPAM) microgels adsorbed to interfaces. The microgels react to changes in temperature by shrinking in aqueous solution above their volume phase transition temperature (VPTT). Additionally, they shrink in mixtures of water and ethanol, although both individual liquids are good solvents for PNIPAM. The combination of this so-called cononsolvency effect and the temperature response of adsorbed microgels is studied by atomic force microscopy (AFM). Adsorbed microgels are of special interest because they are compressed considerably compared to those in bulk solution. It is shown that the impact of adsorption on swelling depends on the specific surface details, as well as the sample preparation. Thereby, the microgels are deposited on two different kinds of surfaces: on gold surface and on polycation (PAH) coating which show different interactions with the microgels in terms of electrostatic interaction and wettability. In addition, the microgels were deposited from different solvent mixtures. This influences the microgel structure and thereby the swelling properties. Nanorheology studies by dynamic AFM measurements lead to surprising results which are explained by the fact that not only polymer density but a subtle interaction between polymer and solvent might dominate the rheological properties. This work supports the view that preferential adsorption of ethanol at PNIPAM drives cononsolvency, while the shrinking at T > VPTT is caused by general breaking of hydrogen bonds between solvents and PNIPAM.

Genome-wide CRISPR-Cas9 screen analyzed by SLIDER identifies network of repressor complexes that regulate TRIM24.

TRIM24 is an oncogenic chromatin reader that is frequently overexpressed in human tumors and associated with poor prognosis. However, TRIM24 is rarely mutated, duplicated, or rearranged in cancer. This raises questions about how TRIM24 is regulated and what changes in its regulation are responsible for its overexpression. Here, we perform a genome-wide CRISPR-Cas9 screen by fluorescence-activated cell sorting (FACS) that nominated 220 negative regulators and elucidated a regulatory network that includes the KAP1 corepressor, CNOT deadenylase, and GID/CTLH E3 ligase. Knocking out required components of these three complexes caused TRIM24 overexpression, confirming their negative regulation of TRIM24. Our findings identify regulators of TRIM24 that nominate previously unexplored contexts for this oncoprotein in biology and disease. These findings were enabled by SLIDER, a new scoring system designed and vetted in our study as a broadly applicable tool for analysis of CRISPR screens performed by FACS.

Mammary-specific expression of Trim24 establishes a mouse model of human metaplastic breast cancer.

Conditional overexpression of histone reader Tripartite motif containing protein 24 (TRIM24) in mouse mammary epithelia (Trim24COE) drives spontaneous development of mammary carcinosarcoma tumors, lacking ER, PR and HER2. Human carcinosarcomas or metaplastic breast cancers (MpBC) are a rare, chemorefractory subclass of triple-negative breast cancers (TNBC). Comparison of Trim24COE metaplastic carcinosarcoma morphology, TRIM24 protein levels and a derived Trim24COE gene signature reveals strong correlation with human MpBC tumors and MpBC patient-derived xenograft (PDX) models. Global and single-cell tumor profiling reveal Met as a direct oncogenic target of TRIM24, leading to aberrant PI3K/mTOR activation. Here, we find that pharmacological inhibition of these pathways in primary Trim24COE tumor cells and TRIM24-PROTAC treatment of MpBC TNBC PDX tumorspheres decreased cellular viability, suggesting potential in therapeutically targeting TRIM24 and its regulated pathways in TRIM24-expressing TNBC.

Structural control of polyelectrolyte/microemulsion droplet complexes (PEMECs) with different polyacrylates.

The ionic assembly of oppositely charged polyelectrolyte-surfactant complexes (PESCs) is often done with the aim of constructing more functional colloids, for instance as advanced delivery systems. However, PESCs are often not easily loaded with a solubilisate due to intrinsic restrictions of such complexes. This question was addressed from a different starting point: by employing microemulsion droplets as heavily loaded surfactant systems and thereby avoiding potential solubilisation limitations from the beginning. We investigated mixtures of cationic oil-in-water (O/W) microemulsion droplets and oppositely charged sodium polyacrylate (NaPA) and determined structure and phase behaviour as a function of the mixing ratio for different droplet sizes and different M w (NaPA). Around an equimolar charge ratio an extended precipitate region is present, which becomes wider for larger droplets and with increasing M w of the NaPA. Static and dynamic light scattering (SLS and DLS) and small-angle neutron scattering (SANS) show the formation of one-dimensional arrangements of microemulsion droplets for polyelectrolyte excess, which become more elongated with increasing M w (NaPA) and less so with increasing NaPA excess. What is interesting is a marked sensitivity to ionic strength, where already a modest increase to ∼20 mM leads to a dissolution of the complexes. This work shows that polyelectrolyte/microemulsion complexes (PEMECs) are structurally very versatile hybrid systems, combining the high solubilisate loading of microemulsions with the larger-scale structuring induced by the polymer, thereby markedly extending the concept of conventional PESCs. This type of system has not been described before and is highly promising for future applications where high payloads are to be formulated.

Poly(N-isopropylacrylamide) Microgels under Alcoholic Intoxication: When a LCST Polymer Shows Swelling with Increasing Temperature.

Poly(N-isopropylacrylamide) (PNIPAM) microgel is a smart polymer that shows a volume phase transition temperature (VPTT) at around 32 °C in aqueous solutions, above which it collapses. In this work, combining experiments and molecular simulations, it is shown that PNIPAM microgels do not always exhibit a collapsed structure above the VPTT. Instead, PNIPAM in aqueous alcohol mixtures shows a two-step conformational transition, i.e., a collapse at low temperatures (T < 32 °C) and a reswelling when T > 50 °C. The present analysis indicates that delicate microscopic interaction details, together with the bulk solution properties, play a key role in dictating the reswelling behavior. Even when PNIPAM microgels swell with increasing T, this is not a standard upper critical solution behavior.

Uptake of pH-Sensitive Gold Nanoparticles in Strong Polyelectrolyte Brushes.

The impact of electrostatic attraction on the uptake of gold nanoparticles (AuNPs) into positively charged strong poly-[2-(Methacryloyloxy) ethyl] trimethylammonium chloride (PMETAC) polyelectrolyte brushes was investigated. In this work, PMETAC brushes were synthesized via surface-initiated atom transfer radical polymerization (Si-ATRP). PMETAC/AuNP composite materials were prepared by incubation of the polymer brush coated samples into 3-mercaptopropionic acid-capped AuNP (5 nm in diameter) suspension. The electrostatic interactions were tuned by changing the surface charge of the AuNPs through variations in pH value, while the charge of the PMETAC brush was not affected. Atomic-force microscopy (AFM), ellipsometry, UV/Vis spectroscopy, gravimetric analysis and transmission electron microscopy (TEM) were employed to study the loading and penetration into the polymer brush. The results show that the number density of attached AuNPs depends on the pH value and increases with increasing pH value. There is also strong evidence that the particle assembly is dependent on the pH value of the AuNP suspension. Incubation of PMETAC brushes in AuNP suspension at pH 4 led to the formation of a surface layer on top of the brush (2D assembly) due to sterical hindrance of the clustered AuNPs, while incubation in AuNP suspension at pH 8 led to deeper particle penetration into the brush (3D assembly). The straightforward control of particle uptake and assembly by tuning the charge density of the nanoparticle surface is a valuable tool for the development of materials for colorimetric sensor applications.