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Background: Combination antiretroviral therapy has significantly advanced HIV-1 infection treatment. However, HIV-1 remains persistent in the brain; the inaccessibility of the blood–brain barrier allows for persistent HIV-1 infections and neuroinflammation. Nanotechnology-based drug carriers such as nanodiscoidal bicelles can provide a solution to combat this challenge. Methods: This study investigated the safety and extended release of a combination antiretroviral therapy drug (tenofovir)-loaded nanodiscs for HIV-1 treatment in the brain both in vitro and in vivo. Result: The nanodiscs entrapped the drug in their interior hydrophobic core and released the payload at the desired location and in a controlled release pattern. The study also included a comparative pharmacokinetic analysis of nanodisc formulations in in vitro and in vivo models. Conclusion: The study provides potential applications of nanodiscs for HIV-1 therapy development.

Fig. S1. CARTmeso cell manufacturing and treatment schedules. Fig. S2. Detection of human anti-chimeric antibody responses in patients post-CARTmeso cell infusion. Fig. S3. Serum cytokines and chemokines in PDA patient 21211-101. Fig. S4. CARTmeso cytolytic activity against allogeneic and autologous pancreatic cancer cell lines Table S1. Patient demographics. Table S2. Treatment-related adverse events. Table S3. Protoarray analysis of serum samples from mesothelioma patient 17510-105 Table S4. Protoarray analysis of serum samples from pancreatic cancer patient 21211-101

Off-target toxicity due to the expression of target antigens in normal tissue represents a major obstacle to the use of chimeric antigen receptor (CAR)-engineered T cells for treatment of solid malignancies. To circumvent this issue, we established a clinical platform for engineering T cells with transient CAR expression by using in vitro transcribed mRNA encoding a CAR that includes both the CD3-ζ and 4-1BB costimulatory domains. We present two case reports from ongoing trials indicating that adoptive transfer of mRNA CAR T cells that target mesothelin (CARTmeso cells) is feasible and safe without overt evidence of off-tumor on-target toxicity against normal tissues. CARTmeso cells persisted transiently within the peripheral blood after intravenous administration and migrated to primary and metastatic tumor sites. Clinical and laboratory evidence of antitumor activity was shown in both patients, and the CARTmeso cells elicited an antitumor immune response revealed by the development of novel antiself antibodies. These data show the potential of using mRNA-engineered T cells to evaluate, in a controlled manner, potential off-tumor on-target toxicities and show that short-lived CAR T cells can induce epitope spreading and mediate antitumor activity in patients with advanced cancer. Thus, these findings support the development of mRNA CAR-based strategies for carcinoma and other solid tumors. Cancer Immunol Res; 2(2); 112–20. ©2013 AACR.

Purpose: Previously, we showed that adoptive transfer of in vivo vaccine-primed and ex vivo (anti-CD3/anti-CD28) costimulated autologous T cells (ex-T) at day +12 after transplant increased CD4 and CD8 T-cell counts at day +42 and augmented vaccine-specific immune responses in patients with myeloma. Here, we investigated the safety and kinetics of T-cell recovery after infusing ex-T at day +2 after transplant.Experimental Design: In this phase I/II two-arm clinical trial, 50 patients with myeloma received autografts after high-dose melphalan followed by infusions of ex-T at day +2 after transplant. Patients also received pretransplant and posttransplant immunizations using a pneumococcal conjugate vaccine only (arm B; n = 24) or the pneumococcal conjugate vaccine plus an HLA-A2–restricted multipeptide vaccine for HLA-A2+ patients (arm A; n = 26).Results: The mean number of T cells infused was 4.26 × 1010 (range, 1.59-5.0). At day 14 after transplant, the median CD3, CD4, and CD8 counts were 4,198, 1,545, and 2,858 cells/μL, respectively. Interleukin (IL)-6 and IL-15 levels increased early after transplant and IL-15 levels correlated significantly to day 14 T-cell counts. Robust vaccine-specific B- and T-cell responses were generated. T-cell infusions were well tolerated with no effect on hematopoietic recovery. Eight patients (16%) developed a T-cell “engraftment syndrome” characterized by diarrhea and fever that was clinically and histopathologically indistinguishable from grade 1 to 3 acute graft-versus-host disease (GVHD) of the gastrointestinal tract (seven patients) and/or grade 1 to 2 cutaneous GVHD (four patients).Conclusions: Adoptive T-cell transfers achieve robust T-cell recovery early after transplant and induce moderate-to-severe autologous GVHD in a subset of patients.

Supplementary Data from Rapid Immune Recovery and Graft-versus-Host Disease–like Engraftment Syndrome following Adoptive Transfer of Costimulated Autologous T Cells

Reciprocal gene set enrichment analysis reveals that SPOP F102C, F133V, and F133L mutants have similar effects on the PC transcriptome (S1); AR Activity Score of PC cells expressing wt or mutant SPOPs (S2); Effects of wt-SPOP and PC-associated SPOP mutants on mRNA expression of AR target genes in Abl PC cells (S3); The transcriptomic response induced by mt-SPOPs in PC cells enriches strongly for target genes of AR antagonists (S4); The impact of SPOP on AR protein levels in Abl cells is post-translational (S5); Wt-SPOP, but not its PC-associated mutants, suppresses AR protein levels in androgen-dependent PC cells (S6); The interaction between SPOP-wt and AR can occur even in the absence of SRC-3 protein (S7); Interaction and binding of SPOP-wt to AR is mediated by an SBC motif (a.a.646-a.a.651) located in the Hinge Region of the AR protein (S8); Endogenous SPOP interacts with and regulates the expression levels of the AR protein in PC cells (S9); The impact of SPOP on AR protein levels in 22Rv1 cells is post-translational (S10); Growth promoting effect of PC-associated SPOP mutants in the context of AR axis inhibition (S11); SPOP gene signature scores in human PC specimens correlate strongly with androgen receptor activity (S12).

Supplementary Figures 1-6 from Multiple Injections of Electroporated Autologous T Cells Expressing a Chimeric Antigen Receptor Mediate Regression of Human Disseminated Tumor

Description of additional methods and procedures used in the study. Also includes Supplemental References.

Legends for Supplemental Figures S1-S12 and Supplemental Table S1.

Supplementary Methods from Multiple Injections of Electroporated Autologous T Cells Expressing a Chimeric Antigen Receptor Mediate Regression of Human Disseminated Tumor

Primers used for PCR cloning and qPCR assays.

Glioblastoma Multiforme (GBM) is a Grade IV malignant brain cancer that is associated with a high recurrence and low survival rate amongst affected patients. Despite advances in the different methods of therapy, the prognosis for GBM has not improved through the years and, thus, alternative treatment methods for GBM are needed. Minocycline (MINO) is known as a semi-synthetic tetracycline that serves as an antibiotic and has also been shown to be able to suppress angiogenesis for GBM treatment. Collected from brown seaweed, alginate is a biodegradable polysaccharide that can be used to form a hydrogel for drug delivery due to its biocompatibility properties. Injectable alginate scaffolds may be a promising component for an adjuvant treatment method against GBM as the injectability property of the scaffold will allow for the site of interest to be filled precisely after tumor resection. The objective of this study is to develop injectable alginate scaffolds for the delivery of MINO for the treatment of GBM. The effects of the concentration of sodium alginate (SA) and calcium carbonate (CaCO3) (i.e., 0.75, 1.00, 1.50, and 2.00 wt./vol.%) with a 0.25 wt./vol.% of glucono-delta lactone (GDL) on the pH, gelation time, dimensions, degradation, and drug release kinetics of alginate scaffolds were investigated. Injectable alginate scaffolds were fabricated by dissolving SA and CaCO3 (1:1) in water, homogenized with GDL for 20 sec., and then injected into a 24-well plate to develop uniform scaffolds. Their properties were evaluated by testing pH values via a pH meter, timing gelation with a digital timer, utilizing a digital microcaliper to investigate dimensions, an analytical scale to evaluate the mass of dried scaffolds to determine scaffold degradation, and evaluating the release rate of MINO using a microplate reader at a wavelength of 350 nm. As the concentration of SA and CaCO3 increased, an increase in pH, gelation time, and overall stability of the scaffold were observed. All four concentrations of SA and CaCO3 tested resulted in scaffolds with desired pH and workable gelation time; however, the 1 wt./vol.% group may be the most ideal as an optimal pH was reached after 10 minutes and the gelation time for these scaffolds was ~8 minutes. Lower concentration scaffolds (0.75 and 1.00 wt./vol.%) degraded faster in comparison to those made with higher concentrations. Drug release kinetics data revealed that the scaffolds can sustain short term release but could be improved by incorporating the drug into micro or nanoparticles before incorporation into the scaffolds. In conclusion, the injectable alginate scaffolds developed in this study may be promising biomaterials for the delivery of drugs for GBM treatment. Citation Format: Kaitlyn D. Ybáñez, Marco A. Arriaga, Rene N. Rico, Theresia Rookstool, Sue Anne Chew. Injectable alginate scaffolds for the delivery of minocycline for the treatment of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2000.

Glioblastoma multiforme (GBM) is the most prevalent and aggressive form of malignant brain tumor and accounts for 49.1% of all central nervous system tumors. Current methods of therapy remain minimally successful as survival rate remains low (only 6.8% of patients surviving 5 years post-diagnosis). Thus, there is a great need to design alternative treatment methods for patients with GBM. Temozolomide (TMZ) is a chemotherapeutic treatment and a first line treatment option for GBM as it has been shown to increase survival time. Minocycline (MINO) is commonly used for its antibiotic properties but has also been proven to have antiangiogenic effects for GBM treatment. The objective of this study was to investigate the additive effects of MINO and TMZ in both GBM (U87) and human umbilical vein endothelial cells (HUVEC) for the treatment of GBM. We hypothesized that the combination of MINO and TMZ will result in lower cell viability and higher protein levels of apoptosis and autophagy markers) compared to the drugs alone and therefore, will be more effective at treating GBM. In this study, U87 and HUVEC cells were treated with MINO and TMZ alone or in combination and a MTT assay was used to determine cell viability by measuring absorbance (570 nm) with a microplate reader. The IC50 values of the drugs were determined from the dose-response curves using GraphPad to compare the sensitivity of each cell line to the drugs. Protein levels of apoptosis (Cas3, PARP) and autophagy (LC3B) markers were determined after treatment of U87 cells with MINO and TMZ alone or in combination using immunoblotting. β-actin was used as a loading control and 0.5% dimethyl sulfoxide was used as a positive autophagy control. Protein bands were detected and visualized with enhanced chemiluminescence reagents. The IC50 of MINO and TMZ were found to be 1688 µM and 1913 µM in U87 cells, and 247 µM and 1162 µM in HUVEC cells, respectively. Although glioma cells result in autophagy and apoptosis when exposed to MINO, MINO’s anti-angiogenic effect may have resulted in heightened sensitivity in HUVEC cells. U87 cells exposed to MINO (1000 µM) resulted in a cell viability of 61% and TMZ (1200 µM) resulted in 60%, while the combination of both drugs resulted in 43% cell viability. HUVEC cells exposed to MINO (200 µM) resulted in a cell viability of 64% and TMZ (30 µM) resulted in 84%, while the combination of both drugs resulted in 44% cell viability, resulting in additive effects for both cell lines. Preliminary results for protein expression analysis show similar expression of apoptotic and autophagy markers in samples with individual and combined drug treatments. In conclusion, this study demonstrated that the combination of MINO and TMZ resulted in additive effects compared to each drug alone and thus, the dual delivery of both drugs from a local delivery system may be a promising therapy for GBM. Citation Format: Marco A. Arriaga, Natasha Garcia-Rodriguez, Andrea Ramirez, Sue Anne Chew. The combination of temozolomide and minocycline for the treatment of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2687.

Ovarian cancer is the fifth leading cause of cancer mortality among women. This high mortality rate is linked to the development of resistance to first line chemotherapy with platinum compounds which has been attributed in part to increased activity of focal adhesion kinase (FAK). The anti-diabetic drug metformin was previously shown to induce cytotoxicity in PROC cells and thus, the combination of a FAK inhibitor, Y15 and metformin may be a promising treatment for PROC. Biomaterial scaffolds can be utilized to deliver drugs locally to maximize the drug concentration and bioactivity at the target site while minimizing non-target systemic toxicity. The Poly(lactic-co-glycolic acid) (PLGA) is a polyester copolymer and its degradation rate can be easily tailored from days to years providing versatility in the delivery of different drugs. The objectives of this study were to investigate the ability of the PLGA scaffold to control the release of the drugs and if the combined delivery of both Y15 and Metformin would result in additive effects on cell viability compared to the release of each drug alone. Scaffolds were fabricated from an easy and economical mold-less technique by combining PLGA and the drugs (i.e. metformin and/or Y15) in tetraglycol and injected into PBS, to form a globular morphology. An MTT assay was used to analyze cell viability in PROC OVCAR3 cells at an absorbance of 570 nm with a microplate reader. Drug loading from the scaffolds were determined using HPLC at absorbance wavelengths of 233 and 376 nm for Metformin and Y15, respectively. Release kinetics were determined using a spectrometer at absorbance wavelengths of 233 and 380 nm for Metformin and Y15 respectively at each timepoint. Metformin and Y15 treatment reduced cell viability by 34 and 46%, respectively, and 76% when combined. In studies with the fabricated PLGA scaffolds, Y15 only and metformin only scaffolds reduced cell viability by 36 and 11% respectively. When the drugs were combined in the scaffolds, there was a 95% reduction in cell viability. Drug loading and loading efficiency for Metformin only and combination scaffolds were 0.577 and 0.584%, respectively and 6.35 and 6.45%, respectively. Drug loading and loading efficiency for Y15 were 0.059 and 0.064%, respectively, and 12.01 and 14.23%, respectively. A lot of drug was lost into the setting solvent during fabrication. This can be improved by optimizing the setting solvent for Metformin and Y15. Only a short term sustained release was observed from the drug encapsulated scaffolds but may be improved by incorporating the drugs into particles prior to loading into the scaffolds. Although drug loading was low, it was still effective at resulting in cell death. In conclusion, the delivery of Y15 and Metformin in a PLGA biomaterial scaffold can result in an additive effect on cell viability and can be further explored as a promising approach for the treatment of PROC. Citation Format: Hannah Obregon, Viviana Villalobos, Manuel A. Duarte, Emily Jordan, Arkene Levy, Sue Anne Chew. The application of a PLGA scaffold for the dual delivery of Y15 and metformin for the treatment of platinum resistant ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2010.

Glioblastoma multiforme (GBM) is the most prevalent and aggressive form of glioma, with poor prognosis and high mortality rates. As GBM is a highly vascularized cancer, antiangiogenic therapies to halt or minimize the rate of tumor growth are critical to improving treatment. In this review, antiangiogenic therapies, including small-molecule drugs, nucleic acids and proteins and peptides, are discussed. The authors further explore biomaterials that have been utilized to increase the bioavailability and bioactivity of antiangiogenic factors for better antitumor responses in GBM. Finally, the authors summarize the current status of biomaterial-based targeting moieties that target endothelial cells in GBM to more efficiently deliver therapeutics to these cells and avoid off-target cell or organ side effects.

The Biomedical Freshman Research Initiative (BFRI) is a course-based undergraduate research experience (CURE) at The University of Texas Rio Grande Valley (UTRGV), a hispanic-serving institution. O...

Immunotherapy is a relatively newer and promising strategy to combat cancer compared with traditional chemotherapy and radiotherapy strategies. The immune responses are often suppressed in the tumor microenvironment and immunotherapy can be applied to activate or enhance the host immune system response to attack and destroy tumor cells. However, immunotherapy applications by themselves are often associated with low response rates; thus, combination therapies are needed to result in effective anticancer treatments. In this chapter, we discuss the nanoscale biomaterial-based strategies, namely, nanoparticles, to deliver combined immunotherapy and chemotherapy, which enable synergistic effect on anticancer therapy. Moreover, we focus the attention on microrobots/nanorobots (MNRs) that have been employed for drug delivery as potential innovative carriers for immunotherapy and chemotherapy. MNRs are promising solutions for drug delivery applications as they can more precisely access parts of the body that are hard to reach without affecting healthy tissues.

ObjectivesTen per cent of patients diagnosed with pancreatic cancer undergo pancreaticoduodenectomy. There is limited previous research focusing on psychological well-being; unmet support needs impact negatively on quality of life. This paper reports the psychological impact of a pancreatic cancer diagnosis and subsequent pancreaticoduodenectomy, exploring how patients’ lives alter following surgery and how they seek support.DesignInductive qualitative study involving in-depth semistructured interviews with 20 participants who had undergone pancreaticoduodenectomy for pancreatic or distal biliary duct cancer. Interviews were audiorecorded, transcribed and anonymised, and thematic analysis used principles of constant comparison.SettingSingle National Health Service Trust in Northwest England.ParticipantsPatients were eligible for inclusion if they had had pancreaticoduodenectomy for head of pancreas cancer, periampullary cancer or distal cholangiocarcinoma between 6 months and 6 years previously, and had completed adjuvant chemotherapy.ResultsAnalysis identified the following main themes: diagnosis and decision making around surgery; recovery from surgery and chemotherapy; burden of monitoring and ongoing symptoms; adjusting to ‘a new normal’; understanding around prognosis; support-seeking. Participants seized the chance to have surgery, often without seeming to absorb the risks or their prognosis. They perceived that they were unable to control their life trajectory and, although they valued close monitoring, experienced anxiety around their appointments. Participants expressed uncertainty about whether they would be able to return to their former activities. There were tensions in their comments about support-seeking, but most felt that emotional support should be offered proactively.ConclusionsPatients should be made aware of potential psychological sequelae, and that treatment completion may trigger the need for more support. Clinical nurse specialists (CNSs) were identified as key members of the team in proactively offering support; further training for CNSs should be encouraged. Understanding patients’ experience of living with cancer and the impact of treatment is crucial in enabling the development of improved support interventions.

Background: The utilization of iron oxide nanoparticles (Fe3O4 NPs) to control minocycline release rates from poly(lactic-co-glycolic acid) scaffolds fabricated from an easy/economical technique is presented. Results & methodology: A larger change in temperature and amount of minocycline released was observed for scaffolds with higher amounts of Fe3O4 NPs, demonstrating that nanoparticle concentration can control heat generation and minocycline release. Temperatures near a polymer’s glass transition temperature can result in the polymer’s chain becoming more mobile and thus increasing drug diffusion out of the scaffold. Elevated temperature and minocycline released from the scaffold can work synergistically to enhance glioblastoma cell death. Conclusion: This study suggests that Fe3O4 NPs are promising materials for controlling minocycline release from polymeric scaffolds by magnetic hyperthermia for the treatment of glioblastoma.

Chimeric antigen receptor (CAR) T cells have demonstrated promising efficacy, particularly in hematologic malignancies. One challenge regarding CAR T cells in solid tumors is the immunosuppressive tumor microenvironment (TME), characterized by high levels of multiple inhibitory factors, including transforming growth factor (TGF)-β. We report results from an in-human phase 1 trial of castration-resistant, prostate cancer-directed CAR T cells armored with a dominant-negative TGF-β receptor (NCT03089203). Primary endpoints were safety and feasibility, while secondary objectives included assessment of CAR T cell distribution, bioactivity and disease response. All prespecified endpoints were met. Eighteen patients enrolled, and 13 subjects received therapy across four dose levels. Five of the 13 patients developed grade ≥2 cytokine release syndrome (CRS), including one patient who experienced a marked clonal CAR T cell expansion,98% reduction in prostate-specific antigen (PSA) and death following grade 4 CRS with concurrent sepsis. Acute increases in inflammatory cytokines correlated with manageable high-grade CRS events. Three additional patients achieved a PSA reduction of ≥30%, with CAR T cell failure accompanied by upregulation of multiple TME-localized inhibitory molecules following adoptive cell transfer. CAR T cell kinetics revealed expansion in blood and tumor trafficking. Thus, clinical application of TGF-β-resistant CAR T cells is feasible and generally safe. Future studies should use superior multipronged approaches against the TME to improve outcomes.

PURPOSE CD19-targeted chimeric antigen receptor (CAR)–modified T cells demonstrate unprecedented responses in B-cell acute lymphoblastic leukemia (B-ALL); however, relapse remains a substantial challenge. Short CAR T-cell persistence contributes to this risk; therefore, strategies to improve persistence are needed. METHODS We conducted a pilot clinical trial of a humanized CD19 CAR T-cell product (huCART19) in children and young adults with relapsed or refractory B-ALL (n = 72) or B-lymphoblastic lymphoma (n = 2), treated in two cohorts: with (retreatment, n = 33) or without (CAR-naive, n = 41) prior CAR exposure. Patients were monitored for toxicity, response, and persistence of huCART19. RESULTS Seventy-four patients 1-29 years of age received huCART19. Cytokine release syndrome developed in 62 (84%) patients and was grade 4 in five (6.8%). Neurologic toxicities were reported in 29 (39%), three (4%) grade 3 or 4, and fully resolved in all cases. The overall response rate at 1 month after infusion was 98% (100% in B-ALL) in the CAR-naive cohort and 64% in the retreatment cohort. At 6 months, the probability of losing huCART19 persistence was 27% (95% CI, 14 to 41) for CAR-naive and 48% (95% CI, 30 to 64) for retreatment patients, whereas the incidence of B-cell recovery was 15% (95% CI, 6 to 28) and 58% (95% CI, 33 to 77), respectively. Relapse-free survival at 12 and 24 months, respectively, was 84% (95% CI, 72 to 97) and 74% (95% CI, 60 to 90) in CAR-naive and 74% (95% CI, 56 to 97) and 58% (95% CI, 37 to 90) in retreatment cohorts. CONCLUSION HuCART19 achieved durable remissions with long-term persistence in children and young adults with relapsed or refractory B-ALL, including after failure of prior CAR T-cell therapy.

Coronavirus Disease 2019 (COVID-19) remains a global health emergency with limited treatment options, lagging vaccine rates and inadequate healthcare resources in the face of an ongoing calamity. The disease is characterized by immune dysregulation and cytokine storm. Cyclosporine A (CSA) is a calcineurin inhibitor that modulates cytokine production and may have direct antiviral properties against coronaviruses. To test whether a short course of treatment was safe in COVID-19 patients, we treated 10 hospitalized, oxygen requiring, non-critically ill patients with CSA at a starting dose of 9mg/kg/day. Five patients experienced adverse events, none were serious, and transaminitis was most common. No subject enrolled in this trial required intensive care unit (ICU)-level care and all patients were discharged alive from the hospital. Further, CSA treatment was associated with significant reductions in serum cytokines and chemokines important in COVID-19 hyper-inflammation, including CXCL10. In conclusion, short courses of CSA appear safe and feasible in COVID-19 patients requiring oxygen and therefore, may be a useful adjunct in resource-poor or resource-limited health care settings.

More than 70 % of women with ovarian cancer are diagnosed with advanced-stage disease, which is initially treated with cytoreductive surgery, and combination chemotherapy with platinum-based compounds. Most patients initially respond to platinum-based therapy, but eventually up to 80 % of this responsive cohort becomes refractory due to the development of platinum resistance. This review discusses current and potential therapeutic approaches that exploit biomaterial-based applications to combat platinum resistance either by enhancing the delivery of platinum-based drugs or prodrugs, delivering other toxic non-platinum-based bioactive factors (by themselves or in combination with platinum-based drugs) or by delivering other bioactive factors that re-sensitize resistant ovarian cancer cells to these drugs. The types of materials that are used, the bioactive factors applied (i.e., drug or gene delivery), and the specific agents that are employed to target these types of cancer cells are discussed. We conclude that the unique attributes of biomaterial-based applications can be further explored toward overcoming platinum-resistant ovarian cancer as monotherapy, or in combination with other treatment strategies.

Glioblastoma multiforme (GBM) is the most common and invasive form of malignant brain tumors and despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with GBM still remains poor. Temozolomide (TMZ) is the chemotherapy drug that is most commonly given orally after surgical resection of these tumors. In this study, the effects of solvents (i.e., dichloromethane and acetonitrile) used for the fabrication of electrosprayed TMZ-loaded poly(lactic-co-glycolic acid) (PLGA) on drug loading, loading efficiency, drug release kinetics, surface morphology, and particle size were investigated. The results from this study demonstrated that by using a larger volume of a solvent with higher polarity (i.e., acetonitrile) which allows for a higher amount of hydrophilic TMZ to dissolve into the polymer solution, higher drug loading could be achieved. However, the particles fabricated with high amount of acetonitrile, which has a lower vapor pressure, had large pores and a smaller diameter which led to an initial burst release and high cumulative release at the end of the study. An optimal combination of the two solvents is needed to result in particles with a good amount of loading and minimal initial burst release. The electrosprayed microparticles were able to illicit a cytotoxic response in U-87 MG glioblastoma cells at a lower concentration of drug compared to the free drug. This work indicated that electrospraying is a promising method for the fabrication of TMZ-loaded PLGA microparticles for the treatment of GBM and solvent composition can be altered to control drug loading and release kinetics. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2317-2324, 2019.

Ovarian cancer is the fifth leading cause of cancer mortality among women in the US. The high mortality rates are linked to the development of resistance to first line chemotherapy with platinum compounds. Currently there is no standard second-line treatment for platinum resistant ovarian cancer (OCpt). Resistance to platinum-based therapy has been attributed in part to increased activity of focal adhesion kinase (FAK). The anti-diabetic drug metformin was previously shown to induce cytotoxicity in OCpt cells and the combination of a FAK inhibitor, Y15 and metformin may be a promising treatment for OCpt. Oftentimes, drugs have a short half-life which results in low bioavailability at the target site of interests. Biomaterial scaffolds can be utilized to deliver drugs locally to maximize the drug concentration and bioactivity at the target site while minimizing non-target systemic exposure and toxicity. Poly(lactic-co-glycolic acid) (PLGA) is an α-hydroxy acid-derived polyester copolymer that has a variety of biomaterial applications due to its great biocompatibility and versatility. The degradation rate of PLGA can be easily tailored from days to years providing versatility in the delivery of different drugs. PLGA scaffolds can be fabricated using a mold-less technique that provides controlled release of drugs from such scaffolds. The objective of this study was to utilize biomaterials as a dual drug delivery system and investigate if the combined delivery of both Y15 and Metformin, would result in synergistic effects on cell viability compared to the release of each drug alone. We also evaluated if controlled release of the drugs could be achieved using the scaffolds. To evaluate this, PLGA scaffolds were fabricated from an easy and economical mold-less technique by combining PLGA and the drugs (i.e. metformin and/or Y15) in tetraglycol and injected in PBS, to form a globular scaffold. An MTT assay was used to analyze cell viability in platinum resistant OVCAR3 ovarian cancer cells at an absorbance of 570 nm with a microplate reader. Drug release from the scaffolds were determined in PBS using a microplate reader at absorbance wavelengths of 233 and 380 nm for Metformin and Y15, respectively. Metformin and Y15 treatment reduced cell viability by 34 and 46%, respectively. In combination, both drugs reduced cell viability by 76%. In studies with the fabricated PLGA scaffolds, Y15 and metformin single treatments reduced cell viability by 36% and 11% respectively. However, when the drugs were combined, there was a 95% reduction in cell viability. Furthermore, the delivery of the drugs from the PLGA scaffolds resulted in a sustained release. In conclusion, the delivery of Y15 and Metformin in a biomaterial scaffold can result in controlled released of the drugs and a synergistic effect on cell viability and thus, can be further explored as promising approach for the treatment of platinum resistant ovarian cancer. Citation Format: Emily M. Jordan, Hannah Obregon, Marco Arriaga, Arkene Levy, Sue Anne Chew. The dual delivery of y15 and metformin in a PLGA scaffold for the treatment of platinum resistant ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1082.

Glioblastoma Multiforme (GBM) is the most frequent and aggressive malignant primary brain tumor in adults with a life expectancy of less than fifteen months after diagnosis. The standard care for GBM includes surgical resection followed by radiotherapy and chemotherapy. Despite advances in the different methods of therapy, the prognosis for gliomas has not been dramatically improved through the years due to high levels of reoccurrence therefore, alternative treatment methods are needed for GBM. Minocycline (MINO) is a common antibiotic with potential anticancer effects by acting as an anti-angiogenic agent which can help in the treatment and reduction of gliomas by reducing blood vessel formation. Temozolomide (TMZ) is a chemotherapy alkylating agent that has demonstrated antitumor activity against highly resistant malignancies as high-grade gliomas. Injectable alginate scaffolds can serve as a local delivery system for controlled drug release and can be an important component for the development of a treatment method against GBM. The objective of this study was to investigate the optimal concentration of TMZ and MINO alone or in combination against glioblastoma cells (U87-MG) and human umbilical vein endothelial cells (HUVEC) and to evaluate the ability of an injectable alginate scaffold to control the rate of drug release for the treatment of GBM. In this study, U87-MG were treated with MINO (0-4000 µM) and TMZ (0-6000 µM), while HUVEC were treated with MINO (0-1000 µM) and TMZ (0-2000 µM) alone or in combination and a MTT assay was used to determine cell viability. Injectable alginate scaffolds were fabricated by dissolving sodium alginate and calcium carbonate in water and then homogenized with glucanolactone and drugs for 20 seconds. The mixture was then injected into a 24-well plate. After fabrication, scaffolds with MINO, TMZ and MINO/TMZ were incubated in PBS at 37 °C. At each timepoint (1, 4, 7, 10 and 14 days), the amount of drug released was measured by reading the absorbance of MINO and TMZ at 350 and 327 nm, respectively using a microplate reader. The 50% inhibitory concentration (IC50) of MINO and TMZ were found to be 950 µM and 900 µM, respectively in U87-MG. In HUVEC, the IC50 of MINO and TMZ were found to be 150 µM and 400 µM, respectively. MINO and TMZ alone reduced cell viability by 39% and 41%, respectively, in U87-MG and in HUVEC by 36% and 34%, respectively. In combination, both drugs reduced cell viability in U87-MG by 58% and in HUVEC by 56%, resulting in synergistic effects for both cell lines. In addition, the application of the injectable alginate scaffold resulted in a controlled release of the drugs alone or in combination. In conclusion, the combination of MINO and TMZ resulted in synergistic effects on cell viability in both cell lines and the controlled delivery of both drugs with an injectable scaffold can be a promising method for the treatment of GBM. Citation Format: Marco A. Arriaga, Rene N. Rico, Kaitlyn D. Ybanez, Daniela Lopez-Lorenzo, Sue Anne Chew. Injectable alginate scaffolds for the dual delivery of Minocycline and Temozolomide for the treatment of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1744.

The adoptive transfer of T lymphocytes reprogrammed to target tumor cells has demonstrated significant potential in various malignancies. However, little is known about the long-term potential and the clonal stability of the infused cells. Here, we studied the longest persisting CD19 redirected chimeric antigen receptor (CAR) T cells to date in two chronic lymphocytic leukemia (CLL) patients who achieved a complete remission in 2010. CAR T-cells were still detectable up to 10+ years post-infusion, with sustained remission in both patients. Surprisingly, a prominent, highly activated CD4+ population developed in both patients during the years post-infusion, dominating the CAR T-cell population at the late time points. This transition was reflected in the stabilization of the clonal make-up of CAR T-cells with a repertoire dominated by few clones. Single cell multi-omics profiling via Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq) with TCR sequencing of CAR T-cells obtained 9.3 years post-infusion demonstrated that these long-persisting CD4+ CAR T-cells exhibited cytotoxic characteristics along with strong evidence of ongoing functional activation and proliferation. Our data provide novel insight into the CAR T-cell characteristics associated with long-term remission in leukemia.

The adoptive transfer of T lymphocytes reprogrammed to target tumour cells has demonstrated potential for treatment of various cancers

Although advances have been made in cancer therapy, cancer remains the second leading cause of death in the U.S. and Europe, and thus efforts to continue to study and discover better treatment methods are ongoing. Three-dimensional (3D) tumor models have shown advantages over bi-dimensional (2D) cultures in evaluating the efficacy of chemotherapy. This commentary aims to highlight the potential of combined application of biomaterials with patient-derived cancer cells as a 3D in vitro model for the study and treatment of cancer patients. Five studies were discussed which demonstrate and provided early evidence to create 3D models with accurate microenvironments that are comparable to in vivo tumors. To date, the use of patient-derived cells for a more personalized approach to healthcare in combination with biomaterials to create a 3D tumor is still relatively new and uncommon for application in clinics. Although highly promising, it is important to acknowledge the current limitations and challenges of developing these innovative in vitro models, including the need for biologists and laboratory technicians to become familiar with biomaterial scaffolds, and the effort for bioengineers to create easy-to-handle scaffolds for routine assessment.

Angiogenesis is an important process in tissue repair and regeneration as blood vessels are integral to supply nutrients to a functioning tissue. In this review, the application of microRNAs (miRNAs) or anti-miRNAs that can induce angiogenesis to aid in blood vessel formation for vascular tissue engineering in ischemic diseases such as peripheral arterial disease and stroke, cardiac diseases, and skin and bone tissue engineering is discussed. Endothelial cells (ECs) form the endothelium of the blood vessel and are recognized as the primary cell type that drives angiogenesis and studied in the applications that were reviewed. Besides ECs, mesenchymal stem cells can also play a pivotal role in these applications, specifically, by secreting growth factors or cytokines for paracrine signaling and/or as constituent cells in the new blood vessel formed. In addition to delivering miRNAs or cells transfected/transduced with miRNAs for angiogenesis and vascular tissue engineering, the utilization of extracellular vesicles (EVs), such as exosomes, microvesicles, and EVs collectively, has been more recently explored. Proangiogenic miRNAs and anti-miRNAs contribute to angiogenesis by targeting the 3'-untranslated region of targets to upregulate proangiogenic factors such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor, and hypoxia-inducible factor-1 and increase the transduction of VEGF signaling through the PI3K/AKT and Ras/Raf/MEK/ERK signaling pathways such as phosphatase and tensin homolog or regulating the signaling of other pathways important for angiogenesis such as the Notch signaling pathway and the pathway to produce nitric oxide. In conclusion, angiogenesis-inducing miRNAs and anti-miRNAs are promising tools for vascular tissue engineering for several applications; however, future work should emphasize optimizing the delivery and usage of these therapies as miRNAs can also be associated with the negative implications of cancer.

CRISPR takes first steps in humans CRISPR-Cas9 is a revolutionary gene-editing technology that offers the potential to treat diseases such as cancer, but the effects of CRISPR in patients are currently unknown. Stadtmauer et al. report a phase 1 clinical trial to assess the safety and feasibility of CRISPR-Cas9 gene editing in three patients with advanced cancer (see the Perspective by Hamilton and Doudna). They removed immune cells called T lymphocytes from patients and used CRISPR-Cas9 to disrupt three genes ( TRAC, TRBC , and PDCD1 ) with the goal of improving antitumor immunity. A cancer-targeting transgene, NY-ESO-1, was also introduced to recognize tumors. The engineered cells were administered to patients and were well tolerated, with durable engraftment observed for the study duration. These encouraging observations pave the way for future trials to study CRISPR-engineered cancer immunotherapies. Science , this issue p. eaba7365 ; see also p. 976

Hepatocellular carcinoma (HCC) is the fifth most common type of cancer diagnosed and the second leading cause of death worldwide. Despite advancement in current treatments for HCC, the prognosis for this cancer is still unfavorable. This comprehensive review article focuses on all the current technology that applies biomaterials to treat and study liver cancer, thus showing the versatility of biomaterials to be used as smart tools in this complex pathologic scenario. Specifically, after introducing the liver anatomy and pathology by focusing on the available treatments for HCC, this review summarizes the current biomaterial-based approaches for systemic delivery and implantable tools for locally administrating bioactive factors and provides a comprehensive discussion of the specific therapies and targeting agents to efficiently deliver those factors. This review also highlights the novel application of biomaterials to study HCC, which includes hydrogels and scaffolds to tissue engineer 3D in vitro models representative of the tumor environment. Such models will serve to better understand the tumor biology and investigate new therapies for HCC. Special focus is given to innovative approaches, e.g., combined delivery therapies, and to alternative approaches—e.g., cell capture—as promising future trends in the application of biomaterials to treat HCC.

Exposure to replication-competent lentivirus (RCL) is a theoretical safety concern for individuals treated with lentiviral gene therapy. For certain ex vivo gene therapy applications, including cancer immunotherapy trials, RCL detection assays are used to screen the vector product as well as the vector-transduced cells. In this study, we reviewed T cell products screened for RCL using methodology developed in the National Gene Vector Biorepository. All trials utilized third-generation lentiviral vectors produced by transient transfection. Samples from 26 clinical trials totaling 460 transduced cell products from 375 subjects were evaluated. All cell products were negative for RCL. A total of 296 of the clinical trial participants were screened for RCL at least 1 month after infusion of the cell product. No research subject has shown evidence of RCL infection. These findings provide further evidence attesting to the safety of third-generation lentiviral vectors and that testing T cell products for RCL does not provide added value to screening the lentiviral vector product.

The p160 steroid receptor coactivators (SRCs) SRC-1, SRC-2 [nuclear receptor coactivator (NCOA)2], and SRC-3 [amplified in breast cancer 1 (AIB1)/NCOA3] are key pleiotropic “master regulators” of transcription factor activity necessary for cancer cell proliferation, survival, metabolism, and metastasis. SRC overexpression and overactivation occur in numerous human cancers and are associated with poor clinical outcomes and resistance to therapy. In prostate cancer (PC), the p160 SRCs play critical roles in androgen receptor transcriptional activity, cell proliferation, and resistance to androgen deprivation therapy. We recently demonstrated that the E3 ubiquitin ligase adaptor speckle-type poxvirus and zinc finger (POZ) domain protein (SPOP) interacts directly with SRC-3 and promotes its cullin 3-dependent ubiquitination and proteolysis in breast cancer, thus functioning as a potential tumor suppressor. Interestingly, somatic heterozygous missense mutations in the SPOP substrate-binding cleft recently were identified in up to 15% of human PCs (making SPOP the gene most commonly affected by nonsynonymous point mutations in PC), but their contribution to PC pathophysiology remains unknown. We now report that PC-associated SPOP mutants cannot interact with SRC-3 protein or promote its ubiquitination and degradation. Our data suggest that wild-type SPOP plays a critical tumor suppressor role in PC cells, promoting the turnover of SRC-3 protein and suppressing androgen receptor transcriptional activity. This tumor suppressor effect is abrogated by the PC-associated SPOP mutations. These studies provide a possible explanation for the role of SPOP mutations in PC, and highlight the potential of SRC-3 as a therapeutic target in PC.

In recent years, the application of microRNAs (miRNAs) or anti-microRNAs (anti-miRNAs) that can induce expression of the runt-related transcription factor 2 (RUNX2), a master regulator of osteogenesis, has been investigated as a promising alternative bone tissue engineering strategy. In this review, biomaterial scaffold-based applications that have been used to deliver cells expressing miRNAs or anti-miRNAs that induce expression of RUNX2 for bone tissue engineering are discussed. An overview of the components of the scaffold-based therapies including the miRNAs/anti-miRNAs, cell types, gene delivery vectors, and scaffolds that have been applied are provided. To date, there have been nine miRNAs/anti-miRNAs (i.e., miRNA-26a, anti-miRNA-31, anti-miRNA-34a, miRNA-135, anti-miRNA-138, anti-miRNA-146a, miRNA-148b, anti-miRNA-221, and anti-miRNA-335) that have been incorporated into scaffold-based bone tissue engineering applications and investigated in an in vivo bone critical-sized defect model. For all of the biomaterial scaffold-based miRNA therapies that have been developed thus far, cells that are transfected or transduced with the miRNA/anti-miRNA are loaded into the scaffolds and implanted at the site of interest instead of locally delivering the miRNA/anti-miRNAs directly from the scaffolds. Thus, future work may focus on developing biomaterial scaffolds to deliver miRNAs or anti-miRNAs into cells in vivo.

Limited cell numbers in umbilical cord blood (UCB) grafts present a major impediment to favorable outcomes in adult transplantation, largely related to delayed or failed engraftment. The advent of UCB transplantation (UCBT) using two grafts successfully circumvents this obstacle, despite the engraftment of only one unit. Preclinical models suggested that the addition of UCB T cells at the time of transplant can enhance engraftment. We tested whether ex vivo activation by CD3/CD28 costimulation and expansion of T cells from a single UCB graft would be safe and feasible in adults with advanced hematologic malignancies, with an overall objective of optimizing engraftment in single unit UCBT. In this phase 1 study, recipients of single UCB units were eligible if the unit was stored in two adequate fractions. Dose limiting toxicity was defined as grade 3 or grade 4 GVHD within 90 days of UCBT. Four patients underwent UCBT; all were treated at the first dose level (10(5) cells/kg). At the 10(5) cells/kg dose level two subjects experienced grade 3 intestinal GVHD, thus meeting stopping criteria. For three subjects, neutrophil engraftment was early (12, 17, and 20 days), while one subject experienced primary graft failure. We observed early donor T cell trafficking and found that expanded T cells produced supraphysiologic levels of cytokines relevant to engraftment and to lymphoid differentiation and function. Taken together, these preliminary data suggest rapid engraftment in recipients of a single UCBT combined with relatively low doses of activated T cells, though potentially complicated by severe GVHD.

In this work, PLGA scaffolds with different architectures were fabricated to investigate the effects of surface area to volume ratio (SVR) (which resulted from the different architectures) on scaffold degradation characteristics and drug release kinetics with minocycline as the model drug. It was hypothesized that the thin strand scaffolds, which had the highest SVR, would degrade faster than the thick strand and globular scaffolds as the increase in surface area will allow more contact between water molecules and degradable ester groups in the polymer. However, it was found that globular scaffolds, which had the lowest SVR, resulted in the fastest degradation which demonstrated that the amount of degradation of the scaffolds does not only depend on the SVR but also on other factors such as the retention of acidic degradation byproducts in the scaffold and scaffold porosity. PLGA 50 : 50 globular scaffolds resulted in a biphasic release profile, with a burst release in the beginning and the middle of the release study which may be beneficial for some drug delivery applications. A clear correlation between SVR and release rates was not observed, indicating that besides the availability of more surface area for drug to diffuse out of the polymer matrix, other factors such as amount of scaffold degradation and scaffold porosity may play a role in determining drug release kinetics. Further studies, such as scanning electron microscopy, need to be performed in the future to further evaluate the porosity, morphology and structure of the scaffolds.