OBJECTIVES:Non-muscle-invasive bladder cancer (NMIBC) refractory to Bacillus Calmette-Guérin (BCG) treatment recurs frequently, even with recent approved therapies. We developed a novel, solvent-free, cancer-specific nanoparticle, PLZ4-coated paclitaxel-loaded micelles (PPM), designed for targeted drug delivery and enhanced efficacy. PLZ4 is a bladder cancer-specific peptide that is decorated on the micelle surface of PPM for cancer-specific drug delivery, while paclitaxel is encapsulated for cytotoxic activity. This first-in-human, single-arm Phase I trial aims to evaluate safety, toxicity, and establish the recommended Phase II dose (RP2D) of intravesical PPM in patients with BCG-unresponsive or intolerant NMIBC. METHODS:Eligible patients have BCG-unresponsive NMIBC with adequate organ function and performance status. Up to 29 participants will be enrolled in the dose-escalation and expansion cohorts. PPM is administered intravesically once weekly for 6 weeks at paclitaxel doses of 25 mg (0.5 mg/mL), 50 mg (1.0 mg/mL), or 75 mg (1.5 mg/mL) in normal 0.9% saline, regardless of patient body size or weight. Primary endpoints are safety and determination of RP2D, using the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE v5.0) for toxicity assessment. Efficacy is evaluated via cystoscopy and urine cytology at 6 weeks post-intervention. PRELIMINARY RESULTS:Three BCG-unresponsive patients have been enrolled and finished treatment at the first dose level. No drug-related adverse event was observed. Two out of these first three patients achieved complete remission. The study is still open for enrollment. CLINICAL TRIAL REGISTRATION:NCT05519241.

CLINICAL TRIAL REGISTRATION:NCT06173349.

While miR-22 is a suppressor of hepatocellular carcinoma (HCC), galectin-1 (Gal-1) serves as a HCC biomarker. Our previous studies have shown the effectiveness of miR-22 gene therapy and silencing Gal-1 as two potential novel options in treating HCC in preclinical mouse models. This study examines the significance of the miR-22-Gal-1 axis in HCC development and treatment. The roles of miR-22 and Gal-1 in human HCC were analyzed using the Cancer Genome Atlas database based on their expression levels. The temporal effects of miR-22 were studied by analyzing signaling pathways affected by miR-22 expression levels during HCC progression. AAV8-miR-22, AAV9-Gal-1 siRNA, and LLS30, a Gal-1 inhibitor, were used to treat orthotopic mouse HCC. Spatial transcriptomics established the location-specific effects of miR-22 in mouse HCC. The signaling pathways affected by miR-22 and Gal-1 were identified by analyzing human HCC transcriptomics compared with those found in miR-22, Gal-1 siRNA, or LLS30-treated mouse HCC. In the early stages of HCC, miR-22-high HCC exhibited extensive upregulation of endobiotic metabolism and xenobiotic detoxification signaling, accompanied by the activation of complement and clotting cascades. In late HCC stages, miR-22-high HCC exhibited heightened innate and adaptive immunity, associated with increased interferon signaling. These impacts were primarily observed in the tumors. At the tumor margin, miR-22 inhibited the Rho GTPase and cell–matrix interaction, revealing its role in reducing matrix remodeling and mobility. In non-tumor areas, miR-22 inhibited inflammation by reducing neutrophil degranulation, platelet activation, chemokine receptor binding, and fiber formation. miR-22, Gal-1 silencing, and LLS30 each exhibited anti-HCC effects and targeted common intracellular signaling pathways. Moreover, the anti-HCC effect of miR-22 was dependent on Gal-1 silencing. miR-22-high/Gal-1-low HCC patients had the best survival outcomes. In addition to the above-mentioned key intracellular pathways, miR-22 gene therapy and Gal-1 siRNA treatment of HCC reduced O-linked glycosylation, suggesting the role of the miR-22-Gal-1 axis in modifying glycosylation, which may affect the extracellular functions of Gal-1. In summary, the miR-22-Gal-1 axis can be an HCC prognostic biomarker, and it has vital roles in regulating metabolism and tumor immunity.

Targeted delivery of antigens and adjuvants to the immune cells without eliciting uncontrolled inflammation is a major challenge in cancer vaccine development. Here, a highly versatile and programmable peptide nucleic acid (PNA)-based vaccine nanoplatform (PVN) is reported to elicit a robust anti-tumor immune response against B16-OVA syngeneic melanoma model. The PVN is built on an 11-mer PNA scaffold, enabling efficient “one-pot” loading of a PNA-modified ovalbumin antigenic peptide (SIINFEKL), CpG adjuvant, and a PNA-derivatized LLP2A ligand (an immune cell and melanoma cell targeting ligand). Super-resolution fluorescence imaging reveals the spatial arrangement of OVA 8 within LP 10-12 [ OVA 8 /CpG/LLP2A ], while circular dichroism spectroscopy confirmsparalleled binding of complementary PNA strands in LP 11 [ OVA 8 /CpG/LLP2A ]. LLP2A displayed on PVNs target activated α4β1 integrin expressed by immune and melanoma cells, boosting antigen presentation by dendritic cells and eliciting strong CD8+T cell   and natural killer cell responses. This amplified antitumor immune response leads to significant tumor regression and prolonged survival of mice bearing syngeneic B16-OVA melanoma. The modular nature and versatility of PVN allow convenient one-pot assembling of peptide antigens, immunomodulators, immune cell and tumor cell targeting ligands, making it practical for the custom design and preparation of personalized cancer vaccines.

The One-Bead One-Compound (OBOC) technology, developed early in my career, has revolutionized combinatorial chemistry and drug discovery. Originating from my work on idiotype-specific peptides, OBOC enables high-throughput screening of diverse libraries on individual beads. Over decades, advancements in decoding, screening, and encoding have expanded its applications to therapeutics, nanomedicine, and functional genomics. This chapter reflects on OBOC's origins, impact, and future potential in biomedical research.

Pancreatic ductal adenocarcinoma (PDA) is a highly aggressive cancer with a 5-year survival rate of approximately 13% in the United States, highlighting the urgent need for effective, clinically translatable therapies beyond those currently offering limited benefit. To address this, we developed a novel small-molecule inhibitor, LLS132, and evaluated its safety and efficacy in multiple preclinical models of PDA. We assessed the antiproliferative activity of LLS132 in vitro using human pancreatic cancer cell lines and organoids derived from the genetically engineered LSL-Kras G12D , LSL-Trp53 R172H/+ , Pdx1-Cre (KPC) mouse model. In vivo efficacy and safety were evaluated using subcutaneous and orthotopic pancreatic allograft models, as well as KPC mice. Pharmacokinetics and tumor penetration were analyzed by LC-MS, while RNA sequencing and western blotting were used to identify molecular pathways affected by LLS132. LLS132 inhibited the growth of human pancreatic cancer cell lines and KPC-derived tumor organoids in vitro. In vivo, LLS132 significantly suppressed tumor growth across all tested models. In a subcutaneous allograft model, LLS132 (30 mg/kg, i.p., 5x/week for 1 week) reduced tumor burden by 79% compared to controls, outperforming gemcitabine (100 mg/kg, i.p., 2x/week), which achieved 48% inhibition. In an orthotopic model, LLS132 at 37.5 mg/kg (75% of the maximum tolerated dose, i.p., 5x/week) reduced tumor volume by 43% after 3 weeks and in the clinically relevant KPC model, LLS132 suppressed tumor progression by 65% over 30 days. Importantly, treatment was well tolerated, with no significant weight loss or liver toxicity observed. Pharmacokinetic analysis revealed a maximum plasma concentration of 19063.7 ± 6656.4 ng/mL at tmax = 1.3± 0.5 hours, and a tumor concentration of 111.5 ± 22.5 ng/mg in KPC mice, confirming effective tumor penetration. Transcriptomic analysis of LLS132-treated Panc-1 cells identified cell cycle regulation as a key pathway, with CDK2 emerging as an important target. LLS132 reduced CDK2 expression in both cancer cells and allograft tumor tissue. In conclusion, LLS132 is a promising therapeutic candidate for PDA, exhibiting potent antitumor activity and favorable safety in preclinical models, mediated in part through suppression of CDK2 and modulation of cell cycle pathways. Irena Krga, Laura Musumeci, Hala Addassi, Kit S. Lam, Ruiwu Liu, Gerardo G. Mackenzie. The novel small molecule inhibitor LLS132 reduces pancreatic cancer growth, in part, through modulation of the cell cycle pathway [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research—Emerging Science Driving Transformative Solutions; Boston, MA; 2025 Sep 28-Oct 1; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2025;85(18_Suppl_3):Abstract nr B078.

Polydiacetylenes (PDAs) possess dramatic chromatic transitions, which can be exploited to make stimuli-responsive sensors. Key to developing robust sensors is understanding the changes in molecular structure responsible for these chromatic transitions. We report systematic multiscale studies of different surfactant PDAs using grazing incidence X-ray diffraction (GIXD), X-ray reflectivity (XRR), and atomic force microscopy (AFM) to determine the structure of PDAs in different phases. Monomer films have significant multilayer registry, which is lost after polymerization to the blue phase. Still the monomer and the blue phase are structurally similar: the molecules are uniformly tilted with the diacetylene motif aligned to enable topochemical polymerization. The transition from the monomer to the blue to red phase PDA is not uniform, and at the molecular scale, all three phases may be present within a film. Measurements with Zn2+ in the subphase or boronic acid functionalized PDA precluded the formation of the red phase, enabling blue phase PDA to be characterized unambiguously. In the blue phase, the molecules are uniformly tilted without registry between multilayers. In comparison, red phase PDA adopts a nonplanar conformation with a decrease in molecular tilt. Red phase GIXD patterns can be fit with two potential conformations, a kinked geometry or a twisted geometry. Finally, AFM and XRR measurements reveal that even relatively uniform appearing PDA films are heterogeneous, from no film (bare) regions to monolayer, bilayer, trilayer, and many multilayers at boundaries.

806 Background: Approximately 75% of patients with non-muscle invasive bladder cancer (NMIBC) treated with transurethral resection (TUR) followed by intravesical Bacillus Calmette-Guérin (BCG) experience cancer recurrence. When BCG-unresponsive, most patients continue to have recurrences even with newly approved therapies and nearly 30% progress to invasive stages. We developed a first-in-class bladder cancer-specific nanotherapeutic, PPM, which is administered intravesically. Our preclinical data has demonstrated PPM can selectively target bladder cancer cells and deliver paclitaxel payload into these cancer cells following intravesical or intravenous administration. Methods: This is a 3+3 first-in-human dose escalation trial. Eligible patients must have pathologically confirmed NMIBC, be unresponsive to intravesical BCG therapy (with or without cytotoxic chemotherapy), possess adequate vital organ function, and consent to cystoscopy with TUR for response evaluation. PPM is administered via intravesical instillation once weekly for six weeks. The trial features three dose levels, with paclitaxel doses of 25 mg, 50 mg, and 75 mg. The primary endpoints include safety and the recommended Phase II dose; secondary endpoints encompass response rate, duration of response, systemic drug absorption, and molecular correlative studies. Patients will be followed at 3 month intervals for 2 years or until disease progression. Results: To date, three patients with pathologically confirmed NMIBC have completed a total of 18 treatments at the first dose level without any PPM-related adverse events: one patient was unresponsive to BCG and intravesical mitomycin therapy, while the other two had BCG-unresponsive disease. Two out of the three patients achieved ongoing complete remission for over six and nine months, respectively. The third patient demonstrated persistent disease, but no progression was noted. Enrollment for Dose Level 2 is currently open. Conclusions: PPM has demonstrated promising clinical activity without toxicity at the first dose level in patients with BCG-unresponsive NMIBC. Data at other dose levels will be presented at the meeting (ClinicalTrials.gov identifier: NCT05519241). Clinical trial information: NCT05519241 .

Ischemic injury causes dynamic damage to the native extracellular matrix (ECM), which plays a key role in tissue homeostasis and regeneration by providing structural support, facilitating force transmission, and transducing key signals to cells. The main approach aimed at repairing injury to ischemic tissues is restoration of vascular function. Due to their potential to form capillary niches, endothelial cells (ECs) are of greatest interest for vascular regeneration. Integrin binding to ECM is crucial for cell anchorage to the surrounding matrix, spreading, migration, and further activation of intracellular signaling pathways. In this study, we proposed to establish an in-situ engineering strategy to remodel the ECM at the ischemic site to guide EC endogenous binding and establish effective EC/ECM interactions to promote revascularization. We designed and constructed a dual-function molecule (LXW7)2-SILY, which is comprised of two functional domains: the first one (LXW7) binds to integrin αvβ3 expressed on ECs, and the second one (SILY) binds to collagen. In vitro, we confirmed (LXW7)2-SILY improved EC adhesion and survival. After in situ injection, (LXW7)2-SILY showed stable retention at the injured area and promoted revascularization, blood perfusion, and tissue regeneration in a mouse hindlimb ischemia model. • A dual-function peptide developed for in-situ engineering native extracellular matrix. • The dual-function peptide specifically anchors endogenous endothelial cells to extracellular matrix. • The dual-function peptide promotes vascularized tissue regeneration.

Psoriasis, an immune-mediated inflammatory skin disorder characterized by a chronically relapsing-remitting course, continues to be primarily managed through topical therapy. While oral administration of tyrosine kinase 2 inhibitors (TYK2i) stands as an effective approach for psoriasis treatment, the potential efficacy of topical application of TYK2i remains unexplored. Herein, the carbomer/alginic acid hydrogel is embedded with borneol (BO) as a new topical carrier of TYK2i for achieving enhanced transdermal permeation and anti-psoriasis efficacy. The hydrogel system, i.e., TYK2i-BO-gel, exhibits significantly improved preventative and therapeutic effects in mice models of psoriasiform dermatitis, as evidenced by phenotypical images, psoriasis severity score index (PSI), histology, immunohistochemical staining, and PCR analysis. Remarkably, TYK2i-BO-gel outperforms conventional topical corticosteroid therapy by significantly preventing psoriatic lesion recurrence as measured by a nearly 50 % reduction in ear thickness changes (p < 0.0001), PSI (p < 0.0001) and epidermal thickness (p < 0.05). Moreover, a strengthened anti-inflammatory effect caused by TYK2i-BO-gel is seen in a human skin explant model, implying its potential application for human patients. With the addition of BO, the TYK2i-BO-gel not only increases skin permeability but also inhibits the expression of antimicrobial peptides in keratinocytes and facilitates the anti-Th17 response of TYK2i with suppressed activation of STAT3. Therefore, this work represents the accessibility and effectiveness of TYK2i-BO-hydrogel as a new topical formulation for antipsoriasis management and shows great potential for clinical application.

Macroporous three-dimensional (3D) framework structured melamine foam-based Enzyme-Linked Immunosorbent Assay (f-ELISA) biosensors were developed for rapid, reliable, sensitive, and on-site detection of trace amount of biomolecules and chemicals. Various ligands can be chemically immobilized onto the melamine foam, which brings in the possibility of working with antibodies, nanobodies, and peptides, respectively, as affinity probes for f-ELISA biosensors with improved stability. Different chemical reagents can be used to modify the foam materials, resulting in varied reactivities with antibodies, nanobodies, and peptides. As a result, the f-ELISA sensors produced from these modified foams exhibit varying levels of sensitivity and performance. This study demonstrated that the chemical reagents used for immobilizing antibodies, nanobodies, and peptides could affect the sensitivities of the f-ELISA sensors, and their storage stabilities under different temperatures varied depending on the sensing probes used, with f-ELISA sensors employing nanobodies as probes exhibiting the highest stability. This study not only showcases the versatility of the f-ELISA system but also opens new avenues for developing cost-effective, portable, and user-friendly diagnostic tools with optimized sensitivity and stability.

Derivatives of the potassium-sparing diuretic amiloride are preferentially cytotoxic toward tumor cells relative to normal cells, and have the capacity to target tumor cell populations resistant to currently employed therapeutic agents. However, a major barrier to clinical translation of the amilorides is their modest cytotoxic potency, with estimated IC 50 values in the high micromolar range. Here we report the synthesis of ten novel amiloride derivatives and the characterization of their cytotoxic potency toward MCF7 (ER/PR-positive), SKBR3 (HER2-positive) and MDA-MB-231 (triple negative) cell line models of breast cancer. Comparisons of derivative structure with cytotoxic potency toward these cell lines underscore the importance of an intact guanidine group, and uncover a strong link between drug-induced cytotoxicity and drug lipophilicity. We demonstrate that our most potent derivative called LLC1 is preferentially cytotoxic toward mouse mammary tumor over normal epithelial organoids, acts in the single digit micromolar range on breast cancer cell line models representing all major subtypes, acts on cell lines that exhibit both transient and sustained resistance to chemotherapeutic agents, but exhibits limited anti-tumor effects in a mouse model of metastatic breast cancer. Nonetheless, our observations offer a roadmap for the future optimization of amiloride-based compounds with preferential cytotoxicity toward breast tumor cells.

TPS4615 Background: Bladder carcinoma is one of the top ten cancers in the US with approximately 75% of patients initially presenting with non-myoinvasive bladder cancer (NMIBC). The current standard of care for NMIBC is transurethral resection (TUR) followed by intravesical instillation, usually with Bacille Calmette-Guérin (BCG). Despite combination treatment, 20-80% of patients' disease recurs and 25% progress to invasive stages. The study drug, PLZ4-coated paclitaxel-loaded micelle (PPM), is a solvent-free injectable nanoscale micelle formulation of paclitaxel (PTX). Preclinical data on bladder cancer-specific targeting ligand named PLZ4 has shown potential to specifically deliver chemotherapeutic micelle into bladder cancer cells, both in vitro and in vivo (Zhang, Urol Oncol 2012). In a toxicokinetic study, female Sprague Dawley rats were given intravesicular injections of PPM once weekly for 6 consecutive weeks. No drug related mortality was reported, and no systematic exposure of PPM was observed in toxicokinetic analysis. The purpose of this phase I trial with PPM is to determine the toxicity and recommended Phase II dose. Methods: This is a single arm, open label, Phase I dose-escalation first-in-human trial with an expansion cohort for the treatment of recurrent or refractory NMIBC with PPM. Eligibility includes BCG unresponsive carcinoma in situ or Ta/T1 urothelial carcinoma. Treatment with pembrolizumab is not an inclusion or exclusion criteria as intravesical taxane has comparable efficacy as intravenous pembrolizumab. Up to 29 patients will be recruited at VA Boston Healthcare system. PPM will be given once weekly for six weeks, 50 ml of solution regardless of patient's size or weight will be administered as intravesical instillation. A 3 + 3 design is adopted for this Phase I trial. There are three dose levels at the dose escalation stage: Dose level I: PTX 25 mg or 0.5 mg/ml; Dose Level II: PTX 50 mg or 1.0 mg/ml; Dose Level III: PTX 75 mg or 1.5 mg/ml. At the expansion cohort, up to 12 patients will be treated with PPM at PTX dose of 50 mg or 1.0 mg/ml to determine the efficacy. The primary objective is to evaluate the safety and tolerability of PPM administered through intravesical instillation and determine the recommended Phase II dose of PPM. The secondary objectives are to evaluate the tumor response at 6 weeks after last treatment, determine systemic absorption within 6 hours after intravesical instillation of PPM and assess progression free survival. The efficacy will be determined by urine cytology and cystoscopy after finishing treatment. Molecular correlative studies will be performed. The toxicity will be assessed by Common Terminology Criteria for Adverse Events v5.0. Patients will be followed at 3 month intervals for 2 years or until disease recurrence or progression. This study is currently open for enrollment. NCT05519241. Clinical trial information: NCT05519241 .

The lack of vascularization associated with deep burns delays the construction of wound beds, increases the risks of infection, and leads to the formation of hypertrophic scars or disfigurement. To address this challenge, we have fabricated a multi-functional pro-angiogenic molecule by grafting integrin αvβ3 ligand LXW7 and collagen-binding peptide (SILY) to a dermatan sulfate (DS) glycosaminoglycan backbone, named LXW7-DS-SILY (LDS), and further employed this to functionalize collagen-based Integra scaffolds. Using a large deep burn wound model in C57/BLK6 mice (8–10 weeks old, 26–32g, n = 39), we demonstrated that LDS-modified collagen-based Integra scaffolds loaded with endothelial cells (ECs) accelerate wound healing rate, re-epithelialization, vascularization, and collagen deposition. Specifically, a 2 cm × 3 cm full-thickness skin burn wound was created 48 h after the burn, and then wounds were treated with four groups of different dressing scaffolds, including Integra + ECs, Integra + LDS, and Integra + LDS + ECs with Integra-only as the control. Digital photos were taken for wound healing measurement on post-treatment days 1, 7, 14, 21, 28, and 35. Post-treatment photos revealed that treatment with the Intgera + LDS + ECs scaffold exhibited a higher wound healing rate in the proliferation phase. Histology results showed significantly increased re-epithelialization, increased collagen deposition, increased thin and mixed collagen fiber content, increased angiogenesis, and shorter wound length within the Integra + LDS + ECs group at Day 35. On Day 14, the Integra + LDS + ECs group showed the same trend. The relative proportions of collagen changed from Day 14 to Day 35 in the Integra + LDS + ECs and Integra + ECs groups demonstrated decreased thick collagen fiber deposition and greater thin and mixed collagen fiber deposition. LDS-modified Integra scaffolds represent a promising novel treatment to accelerate deep burn wound healing, thereby potentially reducing the morbidity associated with open burn wounds. These scaffolds can also potentially reduce the need for autografting and morbidity in patients with already limited areas of harvestable skin.

Supplementary materials and methods

Supplementary Methods from In vivo optical imaging of human lymphoma xenograft using a library-derived peptidomimetic against α4β1 integrin

PDF file - 540K, Protein profiling analysis of HER2+ versus HER2- BCSCs

The methods of OB2C library and LLS compounds synthesis

As a measure of cytotoxic potency, half-maximal inhibitory concentration (IC50) is the concentration at which a drug exerts half of its maximal inhibitory effect against target cells. It can be determined by various methods that require applying additional reagents or lysing the cells. Here, we describe a label-free Sobel-edge-based method, which we name SIC50, for the evaluation of IC50. SIC50 classifies preprocessed phase-contrast images with a state-of-the-art vision transformer and allows for the continuous assessment of IC50 in a faster and more cost-efficient manner. We have validated this method using four drugs and 1,536-well plates and also built a web application. We anticipate that this method will assist in the high-throughput screening of chemical libraries (e.g., small-molecule drugs, small interfering RNA [siRNA], and microRNA and drug discovery).