Cargo Materials Research Articles

Recruitment of autophagy initiator TAX1BP1 advances aggrephagy from cargo collection to sequestration.

Autophagy mediates the degradation of harmful material within lysosomes. In aggrephagy, the pathway mediating the degradation of aggregated, ubiquitinated proteins, this cargo material is collected in larger condensates prior to its sequestration by autophagosomes. In this process, theautophagic cargo receptors SQSTM1/p62 and NBR1 drive cargo condensation, while TAX1BP1, which binds to NBR1, recruits the autophagy machinery to facilitate autophagosome biogenesis at the condensates. The mechanistic basis for the TAX1BP1-mediated switch from cargo collection to its sequestration is unclear. Here we show that TAX1BP1 is not a constitutive component of the condensates. Its recruitment correlates with the induction of autophagosome biogenesis. TAX1BP1 is sufficient to recruit the TBK1 kinase via the SINTBAD adapter. We define the NBR1-TAX1BP1-binding site, which is adjacent to the GABARAP/LC3 interaction site, and demonstrate that the recruitment of TAX1BP1 to cargo mimetics can be enhanced by an increased ubiquitin load. Our study suggests that autophagosome biogenesis is initiated once sufficient cargo is collected in the condensates.

A computer-aided, heterodimer-based “triadic” carrier-free drug delivery platform to mitigate multidrug resistance in lung cancer and enhance efficiency

Co-delivering multiple drugs or circumventing the drug efflux mechanism can significantly decrease multidrug resistance (MDR), a major cause of cancer treatment failure. In this study, we designed and fabricated a universal “three-in-one” self-delivery system for synergistic cancer therapy using a computer-aided strategy. First, we engineered two glutathione (GSH)-responsive heterodimers, ERL-SS-CPT (erlotinib [ERL] linked with camptothecin [CPT] via a disulfide bond [SS]) and CPT-SS-ERI (CPT conjugated with erianin [ERI]), which serve as both cargo and carrier material. Next, molecular dynamics simulations indicated that multiple noncovalent molecular forces, including π-π stacking, hydrogen bonds, hydrophobic interactions, and sulfur bonds, drive the self-assembly process of these heterodimers. We then explored the universality of the heterodimers and developed a “triadic” drug delivery platform comprising 40 variants. Subsequently, we conducted case studies on docetaxel (DTX)-loaded ERL-SS-CPT nanoparticles (denoted as DTX@ERL-SS-CPT NPs) and curcumin (CUR)-loaded ERL-SS-CPT NPs (identified as CUR@CPT-SS-ERI NPs) to comprehensively investigate their self-assembly mechanism, physicochemical properties, storage stability, GSH-responsive drug release, cellular uptake, apoptosis effects, biocompatibility, and cytotoxicity. Both NPs exhibited well-defined spherical structures, high drug loading rates, and excellent storage stability. DTX@ERL-SS-CPT NPs exhibited the strongest cytotoxicity in A549 cells, following the order of DTX@ERL-SS-CPT NPs > ERL-SS-CPT NPs > CPT > DTX > ERL. Conversely, DTX@ERL-SS-CPT NPs showed negligible cytotoxicity in normal human bronchial epithelium cell line (BEAS-2B), indicating good biocompatibility and safety. Similar observations were made for CUR@CPT-SS-ERI NPs regarding biocompatibility and cytotoxicity. Upon endocytosis and encountering intracellular overexpressed GSH, the disulfide-bond linker is cleaved, resulting in the release of the versatile NPs into three parts. The spherical NPs enhance water solubility, reduce the required dosage of free drugs, and increase cellular drug accumulation while suppressing P-glycoprotein (P-gp) expression, leading to apoptosis. This work provides a computer-aided universal strategy—a heterodimer-based “triadic” drug delivery platform—to enhance anticancer efficiency while reducing multidrug resistance.

Abstract 3211: Development of first in class zip code drug conjugate in pancreatic ductal adenocarcinoma

Abstract Pancreatic ductal adenocarcinoma (PC) is the fourth most common cancer-related cause of death in Western countries. It is projected that PC will become the second most prevalent cause of cancer-related mortality by the year 2030. Although chemotherapy may offer certain advantages to those who have been diagnosed with pancreatic cancer, its effectiveness is notably constrained by the emergence of chemoresistance and excessive toxicity. In recent studies, it has been established that circulating tumor DNA (ctDNA) serves as a mechanism for cell-specific genetic transfer. We discovered that transposon sequences (known as Zip Codes, ZC) were responsible for ctDNA cell targeting. Therefore, we investigated the possibility of hijacking these ZC to deliver drugs specifically. We began by determining whether PDCA ZC synthesized chemically could target PDCA cells selectively. To achieve this, ZC sequences were labeled with CY5 for high-throughput microscopy. Results show that numerous PDCA-ZC were specific and efficient at targeting PC cell lines while sparing other cancer cell lines. The AluSc8 (PCZC) sequence was chosen for further investigation. The specificity of PCZC for targeting cells was validated using pancreatic islet cells. Based on these findings, we developed a technique for conjugating PCZC with SN38 via a CL2A linker. The results indicated that PCZC-SN38 was a very effective agent at killing PC cells (IC50: 1 μg), but not other cancer cell lines. The therapeutic efficacy of PCZC-N38 was evaluated in vivo using PDX (patient-derived xenografts, Garcia et al Plos One 2013). Maximum tolerated dose testing (MTD) (dosing from 1 to 21 mg of PCZC-SN38/Kg) failed to reach an MTD. Subsequently, we evaluated the efficacy signal of PCZC-SN38 by administering PCZC-SN38 to animals three times per week for three weeks at concentrations of 3 and 30 mg/kg of DNA (equivalent to 0.05 and 0.5 SN38 mg/kg). We used animals treated with vehicle, PCZC alone, and irinotecan (either once per week (50 mg/kg) or three times per week (0.5 mg/Kg) as a comparison. PCZC-SN38 decreased tumor growth by 41% and 70%, respectively, when compared to the vehicle or PCZC alone. The observed efficacy did not result in a change in body weight. In contrast, every animal administered irinotecan at a dosage of 50 mg/kg died within a week. On the contrary, animals that were given irinotecan at a dosage of 0.5 mg/kg demonstrated a growth inhibition of 60% and exhibited remarkable tolerability. Currently, research is being undertaken to determine the therapeutic efficacy of PCZC-SN38 at higher doses. The results outlined in this study illustrate the preliminary application of PC targeted drug delivery using a PC zip code drug conjugate. The results of our study demonstrate an innovative methodology for targeting PC cells specifically with therapeutic cargo material and supports further preclinical development of PCZC-SN38 for PC. Citation Format: Munnever Cinar, Purnachandra Ganji, Andrey Pisarev, Lourdes Martinez-Medina, Dhana S. Bandi, Pavan K. Puvvula, Steven Chang, Daniel Maneval, Bassel El-Rayes, Leon Bernal-Mizrachi. Development of first in class zip code drug conjugate in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3211.

Development of First in Class DNA-Zip Code Drug Conjugate (ZDC) in Multiple Myeloma

Multiple myeloma is a heterogeneous malignant plasma cell disorder with complex molecular and genetic abnormalities. Significant advances have been made in the treatment of multiple myeloma; however, a large proportion of patients fail to attain a favorable response and relapse. Due to efficacy and toxicity, the development of drug conjugates for multiple myeloma has been limited. Our lab discovered that circulating tumor DNA (ctDNA) serves as a cell-specific mechanism for genetic transmission. Certain retrotransposon sequences within the ctDNA were identified as being crucial for the tropism of ctDNA to target tumor cells (Cinar et al bioRxiv.2022). We therefore query whether these transposons can be exploited for the targeted drug delivery. To do so, chemically synthetize and CY5-labeled multiple retrotransposons DNA sequences (a.k.a Zip Codes, ZC) to perform high through put screening assessing for cell targeting. Our findings showed that the AluSp and MER11c DNA sequences (MM-ZC1 and 2) with unique MM mutations were the most efficient and specific DNA sequences for targeting myeloma cells. Furthermore, Flow cytometry and CYTOF validated the specific MM cell targeting on full bone marrow samples from MM patients. Based on this analysis we used MMZC1 for further drug development. We design a drug conjugation approach for the MMZC and screen various drugs for their killing potential. Two potent MM ZC drug conjugates (ZDC) were identified by high throughput viability screening assay: ZC-DM4, a microtubule inhibitor, and ZC-Carfilzomib, an irreversible proteasome inhibitor. We move forward exploration with ZC-DM4 due to limited carfilzomib-linker drug availability. Experiments cancer cell lines in vitro demonstrated that MMZC-DM4 had a potent killing effect on MM cells (IC50: 3.25 nM) but not on other cancer cell lines. These results were the validated in vivo. Maximum tolerated dosing studies (MTD) demonstrated excellent tolerability in all doses (ranges from 1 to 20 mg of MMZC-DM4/Kg (equivalent to 0.007-0.149 mg of DM4/kg). To look for an efficacy signal of MMZC-DM4, MM xenograft mice were given MMZC-DM4 twice a week for three weeks. Due to limitations in production, we treated the animals biweekly with three distinct low doses of MMZC-DM4: 1.9, 3.75, and 7.5 mg/kg (equivalent to 0.014, 0.028, and 0.056 mg of DM4/kg) and compared their response to mice not treated with MMZC-DM4 and mice treated weekly with bortezomib. MMZC-DM4 at such small doses reduced the tumor by 30% without affecting body weight, WBC, HB, AST, or platelets. In contrast, Bortezomib was associated with 90% growth inhibition at the expense of significant weight loss that required treatment interruptions and thrombocytopenia. Currently, investigations are being conducted to examine the therapeutic efficacy of greater doses of MMZC-DM4. This study described the first-in-class MM targeted medicine delivery employing an MM zic code drug conjugate. Our findings suggest new method for targeted delivery therapeutic cargo material directly to cancer cells that could revolutionize cancer treatment.

P62 and NBR1 functions are dispensable for aggrephagy in mouse ESCs and ESC-derived neurons.

Accumulation of protein aggregates is a hallmark of various neurodegenerative diseases. Selective autophagy mediates the delivery of specific cytoplasmic cargo material into lysosomes for degradation. In aggrephagy, which is the selective autophagy of protein aggregates, the cargo receptors p62 and NBR1 were shown to play important roles in cargo selection. They bind ubiquitinated cargo material via their ubiquitin-associated domains and tether it to autophagic membranes via their LC3-interacting regions. We used mouse embryonic stem cells (ESCs) in combination with genome editing to obtain further insights into the roles of p62 and NBR1 in aggrephagy. Unexpectedly, our data reveal that both ESCs and ESC-derived neurons do not show strong defects in the clearance of protein aggregates upon knockout of p62 or NBR1 and upon mutation of the p62 ubiquitin-associated domain and the LC3-interacting region motif. Taken together, our results show a robust aggregate clearance in ESCs and ESC-derived neurons. Thus, redundancy between the cargo receptors, other factors, and pathways, such as the ubiquitin-proteasome system, may compensate for the loss of function of p62 and NBR1.

Accelerating the circular economy transition process for gateway ports: The case of the Port of Zeebrugge

The transition towards a circular economy (CE) has grown in importance during the last decade, and ports are now being viewed as potentially important contributors to this move towards a more sustainable economy. However, several research questions about adopting CE projects in ports remain unanswered. The importance of clustering and secondary resources for circular loops indicates the importance of proximity to either industrial clusters or urban zones. However, gateway ports (GWP), contrary to industrial ports, metropolitan (or urban) ports and diversified port hubs (which combine an industrial function with the proximity to a large urban area), do not benefit from the vicinity of these secondary resources. Nevertheless, even without access to such secondary resources, any port and port managing body (PMB) needs to prepare for the changing competitive landscape, with new cargo flows, material streams and related businesses linked to the circular economy transition of value chains. Here, GWPs, thanks to their nodal position, still have potential to function as platforms for circular transition initiatives arising in other industries. In the present study, we conceptualize a CE transition process, consisting of six steps, especially relevant to GWPs. We then explore the case of the Port of Zeebrugge (Belgium), shortly before its merger with the Port of Antwerp. We propose that GWPs should rely mainly on their potential to attract ‘new value streams’ as the main driver of their CE strategy. Here, top-down initiatives emanating from internal stakeholders, and likely even external ones, appear to provide pathways to CE success.

Extreme-value analysis of intracellular cargo transport by motor proteins.

Extreme-value analysis (EVA) deals with deviations of data from the median of probability distributions. It has been used for various purposes, such as predicting disasters and analyzing sports records. Herein, we extended the use of EVA to investigate nanoscale runners within cells. Motor proteins such as kinesin and dynein run along microtubules, which are protein-containing rails, to deliver cargo (material needed for cells). While the velocity of these runners is markedly affected by large cargos in viscous intracellular environments, the return-level EVA plots showed that the velocity near no-load conditions disclosed the physical difference between the two runners, resulting from their load (force)-velocity relationships. The concave-up force-velocity relationship of dynein hampered the independent and identically distributed conditions for the retrograde velocity dataset and the abnormality of the return-level plot of EVA. Our findings underscore the importance of EVA for assessing the physical properties of proteins in vivo.

Urgency and Mechanism of Structuring Regional Regulations with the Omnibus Law Method

Structuring regional regulations become necessary as more and more the disclosure of various regulatory problems in the region, both regarding with the amount and material of the load. In an effort to encourage structuring regulations in the regions, this article proposes the use of the omnibus method law in the formation of regional regulations through discussion of urgency and mechanisms the use of omnibus law in the formation of regional regulations. The omnibus law itself is a method or technique in the formation of legislation by establishing a rule to change, revoke, or pass several regulations at once. Method This is worthy of consideration for use in structuring local use regulations overcoming the large number of regional regulations and some of them are problematic in substance. At the local regulatory level, this method is also urgent in order to respond to the Employment Creation Act, which was previously established using the omnibus law method, considering this Law and its implementing regulations require improvements to the regional regulations so that synchronous and harmonious. The mechanism that can be taken is an inventory and analysis of several regional regulations with similar or cognate content to be later formed and compiled into one regional regulation. Various regional regulations whose cargo materials have been combined must be declared revoked and not applies again, in order to make it easier to use compared to just make changes or revoke some articles and insert several new articles as carried out in the Job Creation Law.

Dual-Action Pt(IV) Prodrugs and Targeted Delivery in Metal-Organic Frameworks: Overcoming Cisplatin Resistance and Improving Anticancer Activity

AbstractThe conversion of cisplatin Pt(II) into a Pt(IV) complex is a strategy that can be used to overcome cisplatin resistance and side effects, and this involves the functionalization of the axial positions of cisplatin. Using ligands with bioactivity to target specific cancer cell species could lead to targeted treatment and mitigate severe side effects. Furthermore, enhanced cellular uptake could be used to overcome cisplatin resistance. Anchoring the “dual-acting” prodrug in self-assembled hybrid porous metal-organic frameworks (MOFs) has revealed an even more advanced treatment. The exceptional porosity allows high loading of the drug, and the easily tuned functional groups within the frameworks enable various interactions between the drug and the cargo material to optimize controlled release and prevent premature release. Some MOFs have also demonstrated cytotoxicity against several cancer lines. It is expected that the combination of the prodrug and active MOFs could generate a “triple-acting” anticancer agent for more enhanced cancer treatment. This review discusses the development of cisplatin modified with several prodrugs and its anticancer activity in several cancer cell lines. Furthermore, the review summarizes the involvement of cisplatin derivatives in MOFs, highlighting the slow, controlled, and targeted delivery of the drug and its anticancer activity.

Convergence of secretory, endosomal, and autophagic routes in trans-Golgi-associated lysosomes.

At the trans-Golgi, complex traffic connections exist to the endolysosomal system additional to the main Golgi-to-plasma membrane secretory route. Here, we investigated three hits in a Drosophila screen displaying secretory cargo accumulation in autophagic vesicles: ESCRT-III component Vps20, SNARE-binding Rop, and lysosomal pump subunit VhaPPA1-1. We found that Vps20, Rop, and lysosomal markers localize near the trans-Golgi. Furthermore, we document that the vicinity of the trans-Golgi is the main cellular location for lysosomes and that early, late, and recycling endosomes associate as well with a trans-Golgi-associated degradative compartment where basal microautophagy of secretory cargo and other materials occurs. Disruption of this compartment causes cargo accumulation in our hits, including Munc18 homolog Rop, required with Syx1 and Syx4 for Rab11-mediated endosomal recycling. Finally, besides basal microautophagy, we show that the trans-Golgi-associated degradative compartment contributes to the growth of autophagic vesicles in developmental and starvation-induced macroautophagy. Our results argue that the fly trans-Golgi is the gravitational center of the whole endomembrane system.

Synthesis and characterization of methacrylic acid based amphoteric hydrogels: use as a dual drug delivery system

In this study, methacrylic acid-based containing both anionic and cationic groups in its structure p(Methacrylic acid-co-(3-acrylamidopropyl)trimethylammonium chloride) (p(MAA-co-APTMACl)) and p(Methacrylic acid-co-(2-(acryloyloxy)ethyl)trimethylammonium chloride) (p(MAA-AETAC)) hydrogels were synthesized by redox polymerization technique. The morphology, chemical structure and thermal properties of the synthesized hydrogels were characterized by SEM, FT-IR, and TGA. The swelling characterizations of the synthesized MAA-based hydrogels were carried out in various biological environments. Usability of amphoteric hydrogels containing anionic and cationic groups in their structures as a drug carrier cargo material were investigated for the treatment of two different diseases. For this, two of the selected model drugs were loaded simultaneously on hydrogels and their release properties were examined. The drug release mechanisms were investigated according to the Korsmeyer–Peppas model. Rhodamin 6G and ceftriaxone sodium were loaded to p(MAA-co-APTMACl) hydrogel for dual drug release. In addition, Rhodamin 6G and sulfadiazine were loaded simultaneously on the p(MAA-co-AETAC) hydrogel. According to the release mechanism results obtained for p(MAA-co-AETAC) hydrogels, it was determined that it showed a super-state diffusion mechanism. In addition, antibacterial activities of p(MAA-co-APTMACl) and p(MAA-co-AETAC) hydrogels against gram-negative bacteria (Escherichia coli and Pseudomonas Aeruginosa) and gram-positive bacteria (Staphylococcus Aureus and Bacillus Subtilis) were investigated.

Plasma polymer barrier layers to control the release kinetics from dissolvable microneedle patches

AbstractDissolving microneedles are a noninvasive transdermal drug delivery platform that offers painless needle‐free patient treatment. Current strategies to control the kinetics of drug release from the polymeric microneedle patches consist of adjusting the bulk polymer formulations and/or encapsulating the cargo in controlled delivery vehicles. Instead, in this study, the drug release rates were altered by modifying the surface properties of the microneedles using plasma polymer coatings with tailored properties. The main advantage of this one‐step, solvent‐free, and scalable method is that it can be applied to any type of microneedles regardless of their drug cargo or bulk material. Octadiene plasma polymer films were found to suppress the burst release effect of otherwise rapidly dissolving microneedles and to reduce the dissolution rate by a factor of four compared to uncoated control patches. The approach presented in this study offers an alternative, highly tunable, and customizable strategy for controlling drug release rate from microneedle patches.

Cerebral endothelial cell derived small extracellular vesicles improve cognitive function in aged diabetic rats.

Small extracellular vesicles (sEVs) mediate cell-cell communication by transferring their cargo biological materials into recipient cells. Diabetes mellitus (DM) induces cerebral vascular dysfunction and neurogenesis impairment, which are associated with cognitive decline and an increased risk of developing dementia. Whether the sEVs are involved in DM-induced cerebral vascular disease, is unknown. Therefore, we studied sEVs derived from cerebral endothelial cells (CEC-sEVs) of aged DM rats (DM-CEC-sEVs) and found that DM-CEC-sEVs robustly inhibited neural stem cell (NSC) generation of new neuroblasts and damaged cerebral endothelial function. Treatment of aged DM-rats with CEC-sEVs derived from adult healthy normal rats (N-CEC-sEVs) ameliorated cognitive deficits and improved cerebral vascular function and enhanced neurogenesis. Intravenously administered N-CEC-sEVs crossed the blood brain barrier and were internalized by neural stem cells in the neurogenic region, which were associated with augmentation of miR-1 and –146a and reduction of myeloid differentiation primary response gene 88 and thrombospondin 1 proteins. In addition, uptake of N-CEC-sEVs by the recipient cells was mediated by clathrin and caveolin dependent endocytosis signaling pathways. The present study provides ex vivo and in vivo evidence that DM-CEC-sEVs induce cerebral vascular dysfunction and neurogenesis impairment and that N-CEC-sEVs have a therapeutic effect on improvement of cognitive function by ameliorating dysfunction of cerebral vessels and increasing neurogenesis in aged DM rats, respectively.

JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles

Objective:Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs) have a high propensity for thrombosis, which has been attributed to increased blood counts, endothelial cell (EC) dysfunction, and inflammation. The presence of the JAK2V617F mutation in the ECs of MPN patients has been confirmed, but the consequences of EC involvement by JAK2V617F in the pathogenesis of thrombosis are unclear. Endothelial microparticles (EMPs) released from ECs play an important role in endothelial dysfunction and also in the intercellular exchange of biological signals and information. Several studies have revealed that patients with JAK2V617F and a thrombosis history have increased numbers of MPs in their circulation.Materials and Methods:The current study utilized a lentiviral transduction model of JAK2 wild type (JAK2wt) or JAK2V617F encoding green fluorescent protein (GFP) into human umbilical vein ECs to determine the effect of JAK2V617F on ECs. EC infected with JAK2V617F, JAK2WT, and only-GFP were tested after two days of culture.Results:The proteins of ECs that potentially play a role in the development of thrombosis, including endothelial protein C receptor, thrombomodulin, and tissue factor, were detected by flow cytometry analysis with no statistical significance. Increased annexin-V uptake of JAK2V617F and JAK2wt ECs compared to GFP-alone ECs was detected. The EMP production in the supernatants of the EC culture was investigated. Genotyping of the EMPs revealed the presence of genomic DNA and RNA fragments in EMP cargos. JAK2V617F-positive DNA was detected in EMPs released from JAK2V617F-infected ECs and EMPs were shown to carry the genotype of the cell of origin.Conclusion:JAK2V617F-positive EMPs were shown for the first time in the literature. This novel research provides the first evidence that EMPs might regulate neighboring and distant cells via their cargo materials. Thus, the direct effect of JAK2V617F on ECs and their functions suggests that different mechanisms might play a role in the pathogenesis of thrombosis in MPNs.

Mechanism of Atg9 recruitment by Atg11 in the cytoplasm-to-vacuole targeting pathway

Autophagy is a lysosomal degradation pathway for the removal of damaged and superfluous cytoplasmic material. This is achieved by the sequestration of this cargo material within double-membrane vesicles termed autophagosomes. Autophagosome formation is mediated by the conserved autophagy machinery. In selective autophagy, this machinery including the transmembrane protein Atg9 is recruited to specific cargo material via cargo receptors and the Atg11/FIP200 scaffold protein. The molecular details of the interaction between Atg11 and Atg9 are unclear, and it is still unknown how the recruitment of Atg9 is regulated. Here we employ NMR spectroscopy of the N-terminal disordered domain of Atg9 (Atg9-NTD) to map its interaction with Atg11 revealing that it involves two short peptides both containing a PLF motif. We show that the Atg9-NTD binds to Atg11 with an affinity of about 1 μM and that both PLF motifs contribute to the interaction. Mutation of the PLF motifs abolishes the interaction of the Atg9-NTD with Atg11, reduces the recruitment of Atg9 to the precursor aminopeptidase 1 (prApe1) cargo, and blocks prApe1 transport into the vacuole by the selective autophagy-like cytoplasm-to-vacuole (Cvt) targeting pathway while not affecting bulk autophagy. Our results provide mechanistic insights into the interaction of the Atg11 scaffold with the Atg9 transmembrane protein in selective autophagy and suggest a model where only clustered Atg11 when bound to the prApe1 cargo is able to efficiently recruit Atg9 vesicles.

Designing Natural Polymer-Based Capsules and Spheres for Biomedical Applications-A Review.

Natural polymers, such as polysaccharides and polypeptides, are potential candidates to serve as carriers of biomedical cargo. Natural polymer-based carriers, having a core–shell structural configuration, offer ample scope for introducing multifunctional capabilities and enable the simultaneous encapsulation of cargo materials of different physical and chemical properties for their targeted delivery and sustained and stimuli-responsive release. On the other hand, carriers with a porous matrix structure offer larger surface area and lower density, in order to serve as potential platforms for cell culture and tissue regeneration. This review explores the designing of micro- and nano-metric core–shell capsules and porous spheres, based on various functions. Synthesis approaches, mechanisms of formation, general- and function-specific characteristics, challenges, and future perspectives are discussed. Recent advances in protein-based carriers with a porous matrix structure and different core–shell configurations are also presented in detail.

Horizontal Gene Transfer in Multiple Myeloma and Development of a Non-Viral Gene Delivery System

Multiple myeloma (MM) is a heterogeneous malignant plasma cell disorder with complex molecular and genetic abnormalities. While current therapies have improved outcomes in MM, many patients relapse. Part of the development of the resistance to treatment relates to the genetic heterogeneity of tumors that allows for the development of resistant clones. Our laboratory demonstrated that MM circulating tumor DNA (ctDNA) serves as a mechanism for horizontal gene transfer between cancer cells. Using novel 3-D imaging analysis methodologies, we demonstrated that ctDNA target specifically cancer cells of similar tumor phenotype. Furthermore, using novel in vitro models and whole genome sequencing, we identify certain transposable elements (TE) responsible of allowing DNA insertion into cancer cells. Additionally, our analysis discovered a particular retro-transposon sequence unique of multiple myeloma (MM zip-code, MMZC). After chemically synthesizing the MMZC and using flow cytometry methods, we demonstrated that a MMZC was capable of targeting and internalizing within MM cells after 30 minutes of incubation when compared to a control non-carrying TE sequence. Moreover, after culturing for 24 hours, we observed an increase on the number of cells that have internalized the MMZC. To evaluate genome integration, we ligated the MMZC to a linearized CMV-GFP and a linearized CMV-mCherry fragment prior to adding to MM cell culture. Within 12 hours, live imaging showed expression of GFP and mCherry in MM cells. This results are being validated in single cell sequencing. To evaluate a potential therapeutic potential of MMZC cargo delivery, we ligated MMZC to HSV-Tk-GFP linearized vector and tested the cell specificity targeting and killing effect of MMZC-HSV-Tk-GFP in MM, Pancreatic cancer and colon cancer cell lines. MMZC-HSV-Tk-GFP did not elicit a reduction in cell viability. However, MMZC-HSV-Tk-GFP reduce MM cell viability when ganciclovir was added to the cell culture. The capacity of gene delivery has been validated in-vivo using a MM xenograft mice. PCR evaluating for MMZC-HSV-Tk-GFP after 48 hours of tail injection demonstrated that tumor, but not other organs, amplified HSV-Tk. Furthermore, tumor tissue expressed GFP. To our knowledge this is the first description of the use of humanized non-viral synthetic technology capable of delivering therapeutic cargo material into MM cell. DisclosuresMartinez-Medina: Kodikaz Therapeutic Solutions: Current Employment. El-Rayes: Bayer: Research Funding; Pfizer: Research Funding; Merck: Research Funding; Novartis: Research Funding; Boston biomedical: Research Funding; Bristol Myers Squibb: Research Funding; Astra Zeneca: Consultancy, Research Funding; exelixis: Consultancy; erytotech: Research Funding; dicephera pharmaceutical: Consultancy. Gries: Kodikaz Therapeutic Solutions: Current Employment; Feldan Therapeutics: Consultancy. Bernal-Mizrachi: Winship Cancer Institute of Emory University: Current Employment; Takeda Pharmaceuticals: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Bristol Myers Squibb: Consultancy, Membership on an entity's Board of Directors or advisory committees; Kodikaz Therapeutic Solutions: Consultancy, Current holder of individual stocks in a privately-held company, Patents & Royalties; Bigene: Membership on an entity's Board of Directors or advisory committees.

Analysis of unsaturated soil columns with application to bulk cargo liquefaction in ships

Over the last 40 years, there have been several ship losses carrying granular mineral ores that are believed to have been caused by the sudden shifting of the cargo as a result of liquefaction. It is known that the initial moisture content during loading is important for cargo types which are considered susceptible to liquefaction. To limit the moisture content, the degree of saturation must be below 80% in compaction tests designed to reproduce the energy associated with loading cargo into a ship’s hold. During transport, ships can experience storms causing cyclic loading which leads to densification of the cargo and increases in the degree of saturation. There can also be transient increases in pore pressure and associated reductions in the cargo material’s resistance, which when the ship heels increase the likelihood of the cargo shifting and in the extreme, the ship capsizing. In this paper, a recently developed fully coupled dynamic finite element analysis and constitutive model for unsaturated soil is used to perform a parametric study to explore the capabilities of the numerical code and the mechanics of the problem. The numerical analyses are shown to be capable of running for up to 2500 cycles and simulating both cyclic loading and simultaneous consolidation and drainage, a computationally very demanding task. Outcomes of the analyses are used to discuss the factors influencing cargo stability and vessel instability.

Materiality in the Seam Space: Sketches for a Transitional Port City Dome District

Biomass material volatility generates new opportunities for port-city relationships. Alternative energy markets require specialized port facilities to handle new bulk commodities like biomass. Wood pellets, a type of biomass, present warehousing challenges due to combustion danger. The industrial response to this risk has generated new storage forms for port regions. The return to bulk cargo reintroduces materiality as a focus for port city research, which had generally been regarded as a peripheral concern since the advent of the shipping container. The container had come to represent a borderless, ‘fast capitalism’ throughput model, but research on port ‘accidents’ has complicated this reductive globalization narrative. The programmatic dynamism of wood pellet dome structures suggests new spatially-porous possibilities for an interstitial border space at the port-city interface with material commonalities and hybrid potentials for resilient logistics and civic facilities. In contrast to container cargo unitization, the dome signifies the standardization of the coastal/riparian port environment. Dome structures can help ports plan for the complex challenges of cargo material behaviors and increasing extreme weather events. The article begins with wood pellet materiality to then explore programmatic possibilities that industrial construction technology generates. Conceptually, this joins the proposal of port as ‘seam space’ with port-city resilience planning and the porosity celebrated in recent urbanism literature. Scaling up from wood pellet materiality to an interstitial port-city district, the article contributes to calls for increased attention to materiality as a means to envision new urban agendas.

Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants.

Rapid developments in the field of plant genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) systems necessitate more detailed consideration of the delivery of the CRISPR system into plants. Successful and safe editing of plant genomes is partly based on efficient delivery of the CRISPR system. Along with the use of plasmids and viral vectors as cargo material for genome editing, non-viral vectors have also been considered for delivery purposes. These non-viral vectors can be made of a variety of materials, including inorganic nanoparticles, carbon nanotubes, liposomes, and protein- and peptide-based nanoparticles, as well as nanoscale polymeric materials. They have a decreased immune response, an advantage over viral vectors, and offer additional flexibility in their design, allowing them to be functionalized and targeted to specific sites in a biological system with low cytotoxicity. This review is dedicated to describing the delivery methods of CRISPR system into plants with emphasis on the use of non-viral vectors.