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Supplementary figure 8. Experimental design of mice studies. (A) Flow diagram depicts an experimental strategy for PET imaging with different SST analogs in tumor mice models. (B) Flow diagram depicts an experimental strategy for 177Lu-DOTA-EB-TATE therapy in mice models characterized by high- and low- SSTR2 expression and similar tumor growth rate. (C) Flow diagram depicts an experimental strategy involving treatment with different 177Lu-labeled SST analogs in a mice model characterized by high- SSTR2 expression.

Supplemental Table 1: Analysis of immunostaining of SSTR2 in human tissue samples derived from thyroid cancer patients. Score 0-no staining, Score 1-positive staining in 50% of cells. PTC - papillary thyroid cancer, FTC - follicular thyroid cancer, PDTC - poorly differentiated thyroid cancer, MTC - medullary thyroid cancer, HTC - Hürthle cell thyroid cancer.

Supplemental figure 4: VAC treatment did not improve the uptake of SST analogs in the tumors of the mice models. (A) Representative PET images comparing the uptake of 86Y-DOTA-EB-TATE between the control and VAC treated FTC133 subcutaneous mice model. The bar graph shows no significant difference in the uptake of 68Ga-DOTA-TATE, 68Ga-DOTA-JR11 and 86Y-DOTA-EB-TATE between the control (n=5) and VAC (n=5) treated mice. The representative immunohistochemistry images show an increased cytoplasmic expression of SSTR2 in the tumors of the VAC treated mice in comparison to the tumors of the control mice (Supplemental Table 2). (B) Representative PET images comparing the uptake of 86Y-DOTA-EB-TATE between the control and VAC treated FTC133 metastatic mice model. The bar graph shows no significant difference in the uptake of 68Ga-DOTA-TATE, 68Ga-DOTA-JR11 and 86Y-DOTA-EB-TATE between the control (n=8-12) and VAC (n=8-12) treated mice. The immunohistochemistry images show no difference in the expression of SSTR2 in the metastatic tumors of the control and VAC treated FTC133 mice (Supplemental Table 2). (C) Representative PET images comparing the uptake of 86Y-DOTA-EB-TATE between the control and VAC treated TT subcutaneous mice model. The bar graph shows no significant difference in the uptake of 68Ga-DOTA-TATE, 68Ga-DOTA-JR11 and 86Y-DOTA-EB-TATE between the control (n=5) and VAC (n=5) treated mice. The representative immunohistochemistry images show no difference in the expression of SSTR2 within the tumors of the control and VAC treated mice (Supplemental Table 2). (D) Representative PET images comparing the uptake of 86Y-DOTA-EB-TATE between the control and VAC treated AR42J subcutaneous mice model. The bar graph shows no significant difference in the uptake of 68Ga-DOTA-TATE, 68Ga-DOTA-JR11 and 86Y-DOTA-EB-TATE between the control (n=4) and VAC (n=5) treated mice. The representative immunohistochemistry images show an increased expression of SSTR2 in the tumors of the VAC treated mice in comparison to the tumors of the control mice (Supplemental Table 2). Tumors (Tu) and metastasis (Met) are indicated by white arrows. The SUV scales range from 0 to 3 for 68Ga-DOTA-TATE and 68Ga-DOTA-JR11 and 0 to 15 for 86Y-DOTA-EB-TATE. Data are presented as mean{plus minus}SD.

Supplemental Figure 7: VAC treatment did not improve therapeutic efficacy of DOTA-EB-TATE in either low- or high-SSTR2 expressing tumors. (A) No effect of VAC on the tumor progression in the 177Lu-DOTA-EB-TATE treated FTC133 (low-SSTR2-expressing) subcutaneous xenograft mice model. C-control mice that received 177Lu-DOTA-EB-TATE (n=8); VAC-valproic acid treated mice that received 177Lu-DOTA-EB-TATE (n=9). Data are presented as mean{plus minus}SEM. (B) No effect of VAC on the tumor progression in the 177Lu-DOTA-EB-TATE treated AR42J (high-SSTR2-expressing) subcutaneous xenograft mice model. C-control mice that received 177Lu-DOTA-EB-TATE (n=8); VAC-valproic acid treated mice that received 177Lu-DOTA-EB-TATE (n=7). Data are presented as mean{plus minus}SEM.

Supplemental figure 6: Treatment with epigenetic modulators did not increase 86Y-DOTA-EB-TATE uptake within the tumor and normal tissues of the mice models. (A) The bar graph shows quantification of 86Y-DOTA-EB-TATE in the tumor and normal tissues (liver, kidneys, spleen, heart, and lungs) of the control (n=5) and VAC treated (n=5) FTC133 subcutaneous xenograft mice. p=NS (non-significant) w.r.t corresponding control mice tissues; ***p

Supplemental figure 2: Valproic acid is a superior epigenetic modifier in upregulating SSTR2 expression in thyroid cancer cell lines. (A, B) VAC (300 ug/mL, equivalent to 2 mM), TAC (500 ng/mL), and DEC (75 ng/mL) treatment upregulates SSTR2 expression in FTC133 (n=3) and BCPAP (n=3) cells. ***p

Supplemental Table 3: Analysis of immunostaining of SSTR2 in tumor-derived from mice xenografts. Score 0-no staining, Score 1-positive staining in 50% of cells.

Supplemental figure 1: SSTR2 is overexpressed in thyroid cancer tissues in comparison to normal thyroid tissues. (A) SSTR2 expression in different stages of PTC. (B) SSTR2 expression in histological subtypes of PTC. (C) Comparison of SSTR2 mRNA expression in different TC cell lines. (D) TT cells had significantly higher expression than BCPAP and FTC133 cells at the mRNA level. *p

Supplemental Table 2. Analysis of the metastatic lesions characterized by the highest 68Ga-DOTATATE maximum standard uptake values (SUVmax) in 25 patients enrolled in the clinical trial. The data on patients with SUVmax in metastatic lesions exceeding 15 are bolded. PTC - papillary thyroid cancer, PTCTC - tall cell variant PTC, HTC - Hurthle cell thyroid cancer, MTC - medullary thyroid cancer. * uptake similar to background uptake

Purpose:The goal of this study was to analyze the role of somatostatin receptor type 2 (SSTR2) as a molecular target for the imaging and treatment of thyroid cancer through analysis of SSTR2 expression and its epigenetic modulation and testing tumor uptake of different radiolabeled SSTR2 analogues.Experimental Design:We analyzed SSTR2 expression by immunostaining of 92 thyroid cancer tissue samples and quantified standard uptake values (SUVmax) of SSTR2 analogue, 68Ga-DOTA-TATE, by PET/CT imaging in 25 patients with metastatic thyroid cancer. We utilized human thyroid cancer cell lines characterized by differential SSTR2 expression (TT, BCPAP, and FTC133) and rat pancreatic cell line (AR42J) with intrinsically high SSTR2 expression for functional in vitro studies. SSTR2-high (AR42J) and SSTR2-low (FTC133) xenograft mouse models were used to test the uptake of radiolabeled SSTR2 analogues and their therapeutic efficacy in vivo.Results:Thyroid cancer had a higher SSTR2 expression than normal thyroid. Hurthle cell thyroid cancer was characterized by the highest 68Ga-DOTA-TATE uptake [median SUVmax, 16.5 (7.9–29)] than other types of thyroid cancers. In vivo studies demonstrated that radiolabeled DOTA-EB-TATE is characterized by significantly higher tumor uptake than DOTA-TATE (P < 0.001) and DOTA-JR11 (P < 0.001). Treatment with 177Lu-DOTA-EB-TATE extended survival and reduced tumor size in a mouse model characterized by high somatostatin (SST) analogues uptake (SUVmax, 15.16 ± 4.34), but had no effects in a model with low SST analogues uptake (SUVmax, 4.8 ± 0.27).Conclusions:A novel SST analogue, 177Lu-DOTA-EB-TATE, has the potential to be translated from bench to bedside for the targeted therapy of patients characterized by high uptake of SST analogues in metastatic lesions.

Supplemental figure 3: VAC inhibits cell proliferation by promoting cell cycle arrest and apoptosis. (A, D, G, J) VAC treatment (2 mM and 4 mM) inhibits cellular proliferation of FTC133 (n=3), BCPAP (n=3), TT (n=3) and AR42J (n=3) cells. ***p

Supplemental figure 5: Decitabine (DEC) treatment did not improve the uptake of 86Y-DOTA-EB-TATE in the tumors of the mice models. (A) Representative PET images comparing the uptake of 86Y-DOTA-EB-TATE between the control and decitabine (DEC) treated FTC133 subcutaneous mice model. The bar graph shows no significant difference in the uptake of 86Y-DOTA-EB-TATE between the control (n=3) and DEC (n=3) treated mice. (B) Representative PET images comparing the uptake of 86Y-DOTA-EB-TATE between the control and DEC treated AR42J subcutaneous mice model. The bar graph shows no significant difference in the uptake of 86Y-DOTA-EB-TATE between the control (n=5) and DEC (n=3) treated mice. Tumors (Tu) are indicated by white arrows.

The expanded pedicled deltopectoral flap (EPDF) has been widely used to repair large facial scars. Although doctors and patients are usually satisfied with the outcomes, the actual functional recovery and cosmetic effects of EPDF are still unknown. It is, therefore, necessary to objectively investigate the effect of transferred EPDF by using a variety of methods. From January 2008 to December 2018, 52 patients who underwent EPDF surgery at Xijing Hospital were enrolled. Sense of touch, static 2-point discrimination, elasticity, and color were measured. Thermesthesia and algesthesia were also tested. Postoperative scars were evaluated using the patient and observer scar assessment scale. Satisfaction of patients, doctors, and laypersons was investigated. The algaesthesis, thalposis, and rhigosis scores were 4.7 ± 0.7, 3.7 ± 0.9, and 4.5 ± 0.8, respectively. The tactile score was 0.3 ± 0.2 mN, and 2-point discrimination was 10.1 ± 4.8 mm. L*, a*, hemoglobin, and melanin content of the flaps were significantly different when compared with normal skin (P

Purpose: The goal of this study was to analyze the role of somatostatin receptor type 2 (SSTR2) as a molecular target for the imaging and treatment of thyroid cancer through analysis of SSTR2 expression and its epigenetic modulation and testing tumor uptake of different radiolabeled SSTR2 analogues. Experimental Design: We analyzed SSTR2 expression by immunostaining of 92 thyroid cancer tissue samples and quantified standard uptake values (SUVmax) of SSTR2 analogue, 68Ga-DOTA-TATE, by PET/CT imaging in 25 patients with metastatic thyroid cancer. We utilized human thyroid cancer cell lines characterized by differential SSTR2 expression (TT, BCPAP, and FTC133) and rat pancreatic cell line (AR42J) with intrinsically high SSTR2 expression for functional in vitro studies. SSTR2-high (AR42J) and SSTR2-low (FTC133) xenograft mouse models were used to test the uptake of radiolabeled SSTR2 analogues and their therapeutic efficacy in vivo. Results: Thyroid cancer had a higher SSTR2 expression than normal thyroid. Hurthle cell thyroid cancer was characterized by the highest 68Ga-DOTA-TATE uptake [median SUVmax, 16.5 (7.9–29)] than other types of thyroid cancers. In vivo studies demonstrated that radiolabeled DOTA-EB-TATE is characterized by significantly higher tumor uptake than DOTA-TATE (P < 0.001) and DOTA-JR11 (P < 0.001). Treatment with 177Lu-DOTA-EB-TATE extended survival and reduced tumor size in a mouse model characterized by high somatostatin (SST) analogues uptake (SUVmax, 15.16 ± 4.34), but had no effects in a model with low SST analogues uptake (SUVmax, 4.8 ± 0.27). Conclusions: A novel SST analogue, 177Lu-DOTA-EB-TATE, has the potential to be translated from bench to bedside for the targeted therapy of patients characterized by high uptake of SST analogues in metastatic lesions.

Background: Integrins are cell adhesion receptors consisting of 18α and 8β subunits. A subset of integrins (αvβ3) recognizes Arg-Gly-Asp (RGD) peptide motifs which are involved in the neovascularization and progression of various cancers. The aim of the study was to investigate if αvβ3 can serve as a molecular target for the treatment of thyroid cancer (TC) with a novel radiolabeled RGD analog 177Lu-EB-RGD. Methods: Integrin αvβ3 mRNA and/or protein expression was evaluated in 496 TC included in The Cancer Genome Atlas, tissue microarray including 70 TC and 10 normal thyroid samples, and 14 TC cell lines. BRAF-like or RAS-like expression profile was determined through standard BRS scores ranging from -1 to 0 for BRAF-like and 0 to 1 for RAS-like TC. The association between BRS and the αvβ3 expression was tested by the Spearman correlation (r). Nude mice xenografts developing αvβ3 expressing TC after subcutaneous injection of 5*10^6 TC cells were subjected to monotherapy with 0.5 mCi 177Lu-EB-RGD (177Lu) or in combination with Lenvatinib (177Lu+L). The therapeutic efficacy of 177Lu-EB-RGD was compared with standard-of-care Lenvatinib alone (L) and placebo (P). The continuous data were presented as medians with [25-75% interquartile ranges] and compared using Kruskal-Wallis test. The mixed-effects models were used for longitudinal data analysis with adjusted p≤ 0.05 considered statistically significant. Results: We found a moderate negative correlation between BRS and αv (r=-0.5, p Conclusions: The radiolabeled αvβ3 analog 177Lu-EB-RGD has potent growth inhibitory effects in TC characterized by a high integrins expression. The αvβ3 integrin could potentially serve as a molecular target for therapy with radiolabeled RGD analogs in TC. Citation Format: Sonam Kumari, Zhantong Wang, Shilpa Thakur, Laura Abaandou, Oksana Gavrilova, Huiyan Lu, Lixin Lang, Dale Kiesewetter, Vasyl Vasko, Joanna Klubo-Gwiezdzinska. Radiolabeled αvβ3 analog 177Lu-EB-RGD is an effective therapeutic agent in thyroid cancer xenograft mouse model. [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 3946.

It is widely accepted that a small particle size and rough surface can enhance tumor tissue accumulation and tumor cellular uptake of nanoparticles, respectively. Herein, sub-50 nm urchin-inspired disulfide bond-bridged mesoporous organosilica nanoparticles (UMONs) featured with a spiky surface and glutathione (GSH)-responsive biodegradability were successfully synthesized by a facile one-pot biphasic synthesis strategy for enhanced cellular internalization and tumor accumulation. l-Arginine (LA) is encapsulated into the mesopores of UMONs, whose outer surface is capped with the gatekeeper of ultrasmall gold nanoparticles, i.e., UMONs-LA-Au. On the one hand, the mild acidity-activated uncapping of ultrasmall gold can realize a tumor microenvironment (TME)-responsive release of LA. On the other hand, the unique natural glucose oxidase (GOx)-mimicking catalytic activity of ultrasmall gold can catalyze the decomposition of intratumoral glucose to produce acidic hydrogen peroxide (H2O2) and gluconic acid. Remarkably, these products can not only further facilitate the release of LA, but also catalyze the LA-H2O2 reaction for an increased nitric oxide (NO) yield, which realizes synergistic catalysis-enhanced NO gas therapy for tumor eradication. The judiciously fabricated UMONs-LA-Au present a paradigm of TME-responsive nanoplatforms for both enhanced cellular uptake and tumor-specific precision cascaded therapy, which broadens the range of practical biomedical applications and holds a significant promise for the clinical translation of silica-based nanotheranostics.

Zwitterionic surface modification is a promising strategy for nanomedicines to achieve prolonged circulation time and thus effective tumor accumulation. However, zwitterion modified nanoparticles suffer from reduced cellular internalization efficiency. Methods: A polyprodrug-based nanomedicine with zwitterionic-to-cationic charge conversion ability (denoted as ZTC-NMs) was developed for enhanced chemotherapeutic drug delivery. The polyprodrug consists of pH-responsive poly(carboxybetaine)-like zwitterionic segment and glutathione-responsive camptothecin prodrug segment. Results: The ZTC-NMs combine the advantages of zwitterionic surface and polyprodrug. Compared with conventional zwitterionic surface, the ZTC-NMs can respond to tumor microenvironment and realize ZTC surface charge conversion, thus improve cellular internalization efficiency of the nanomedicines. Conclusions: This ZTC method offers a strategy to promote the drug delivery efficiency and therapeutic efficacy, which is promising for the development of cancer nanomedicines.

Combination therapy that could better balance immune activation and suppressive signals holds great potential in cancer immunotherapy. Herein, we serendipitously found that the pH-responsive nanovesicles (pRNVs) self-assembled from block copolymer polyethylene glycol-b-cationic polypeptide can not only serve as a nanocarrier but also cause immunogenic cell death (ICD) through preapoptotic exposure of calreticulin. After coencapsulation of a photosensitizer, 2-(1-hexyloxyethyl)-2-devinyl pyropheophorbide-a (HPPH) and an indoleamine 2,3-dioxygenase inhibitor, indoximod (IND), pRNVs/HPPH/IND at a single low dose elicited significant antitumor efficacy and abscopal effect following laser irradiation in a B16F10 melanoma tumor model. Treatment efficacy attributes to three key factors: (i) singlet oxygen generation by HPPH-mediated photodynamic therapy (PDT); (ii) increased dendritic cell (DC) recruitment and immune response provocation after ICD induced by pRNVs and PDT; and (iii) tumor microenvironment modulation by IND via enhancing P-S6K phosphorylation for CD8+ T cell development. This study exploited the nanocarrier to induce ICD for the host's immunity activation. The "all-in-one" smart nanovesicles allow the design of multifunctional materials to strengthen cancer immunotherapy efficacy.

Dynamic regulation of nanoparticles in a controllable manner has great potential in various areas. Compared to the individual nanoparticles, the assembled nanoparticles exhibit superior properties and functions, which can be applied to achieve desirable performances. Here, a pH-responsive i-motif DNA-mediated strategy to tailor the programmable behaviors of erbium-based rare-earth nanoparticles (ErNPs) decorated copper doped metal-organic framework (CPM) nanohybrids (ECPM) under physiological conditions is reported. Within the acidic tumor microenvironment, the i-motif DNA strands are able to form quadruplex structures, resulting in the assembly of nanohybrids and selective tumor accumulation, which further amplify the ErNPs downconversion emission (1550 nm) signal for imaging. Meanwhile, the ECPM matrix acts as a near-infrared (NIR) photon-activated reactive oxygen species (ROS) amplifier through the singlet oxygen generation of the matrix in combination with its ability of intracellular glutathione depletion upon irradiation. In short, this work displays a classical example of engineering of nanoparticles, which will manifest the importance of developing nanohybrids with structural programmability in biomedical applications.

Background Integrins are cell adhesion receptors consisting of 24 transmembrane heterodimers generated from a combination of 18α integrin and 8β integrin subunits. A subset of integrins consists of receptors recognizing Arg-Gly-Asp (RGD) peptide motifs. One of the RGD-recognizing receptors is integrin αvβ3 that has been recently shown to play a role in neovascularization and progression of several cancers. Radiolabeled RGD analogs have emerged as potential imaging and therapeutic options in cancers characterized by a high expression of integrin αvβ3. Therefore, the aim of this study was to establish the expression of integrin αvβ3 in thyroid cancer (TC). Methods We analyzed the mRNA expression of integrin αvβ3 in 496 BRAF-like and RAS-like human TC tissue samples, including 65 paired samples of tumor vs normal tissue based on The Cancer Genome Atlas. We assessed the protein expression of integrin αvβ3 in 70 TC tissue samples and 10 normal thyroid tissues, as well as in 14 TC cell lines. BRAF-like or RAS-like tumor status was determined by BRS score based upon standard expression profiles ranging from -1 to 0 for BRAF-like cancer and 0 to 1 for RAS-like tumors. The association between BRS and αvβ3 expression was tested using the Spearman correlation coefficient (r). T-tests and paired T-tests were used to compare the continuous variables between the two groups as appropriate, and Kruskal-Wallis test was used for multiple group comparisons with an adjusted p-value of ≤ 0.05 as statistically significant. Results αv integrin subunit mRNA expression was significantly higher in TC than normal thyroid (log fold change 0.3, p=0.001), while the expression of the β3 subunit was similar between paired normal and malignant samples (log fold change -0.2, p=0.30). BRAF-like tumors were characterized by a higher mRNA expression of αvβ3 integrins as documented by a moderate negative correlation between BRS and αv (r=-0.5, p Conclusions TC is characterized by a differential expression of αvβ3 integrin, which is particularly high in the most common type of TC - BRAF-like papillary TC. The αvβ3 integrin could potentially serve as a molecular target for imaging and therapy with radiolabeled RGD analogs in TC. Presentation: Saturday, June 11, 2022 12:00 p.m. - 12:15 p.m.

Reactive oxygen species (ROS) can be used not only as a therapeutic agent for chemodynamic therapy (CDT), but also as a stimulus to activate release of antitumor drugs, achieving enhanced efficacy through the combination of CDT and chemotherapy. Here we report a pH/ROS dual-responsive nanomedicine consisting of β-lapachone (Lap), a pH-responsive polymer, and a ROS-responsive polyprodrug. In the intracellular acidic environment, the nanomedicine can realize pH-triggered disassembly. The released Lap can efficiently generate hydrogen peroxide, which will be further converted into highly toxic hydroxyl radicals via the Fenton reaction. Subsequently, through ROS-induced cleavage of thioketal linker, doxorubicin is released from the polyprodrug. In vivo results indicate that the cascade of ROS generation and antitumor-drug release can effectively inhibit tumor growth. This design of nanomedicine with cascade reactions offers a promising strategy to enhance antitumor efficacy.

Phototheranostics refers to advanced photonics-mediated theranostic methods for cancer and includes imaging-guided photothermal/chemotherapy, photothermal/photodynamic therapy, and photodynamic/chemotherapy, which are expected to provide a paradigm of modern precision medicine. In this regard, various phototheranostic drug delivery systems with excellent photonic performance, controlled drug delivery/release, and precise photoimaging guidance have been developed. In this study, we reported a special "in situ framework growth" method to synthesize novel phototheranostic hollow mesoporous nanoparticles by ingenious hybridization of perylene diimide (PDI) within the framework of small-sized hollow mesoporous organosilica (HMO). The marriage of PDI and HMO endowed the phototheranostic silica nanoparticles (HMPDINs) with largely amplified fluorescence and photoacoustic signals, which can be used for enhanced fluorescence and photoacoustic imaging. The organosilica shell can be chemically chelated with isotope 64Cu for positron emission tomography imaging. Moreover, in situ polymer growth was introduced in the hollow structure of the HMPDINs to produce thermosensitive polymer (TP) in the cavity of HMPDINs to increase the loading capacity and prevent unexpected leakage of the hydrophobic drug SN38. Furthermore, the framework-hybridized PDI generated heat under near-infrared laser irradiation to trigger the deformation of TP for controlled drug release in the tumor region. The fabricated hybrid nanomedicine with organic-inorganic characteristic not only increases the cancer theranostic efficacy but also offers an attractive solution for designing powerful theranostic platforms.

Nanomedicine has become a promising approach to improve cancer chemotherapy. It remains a major challenge how to enhance anti-drug efficacy and reduce side effects of anti-cancer drugs. Herein, we report a self-assembled nanoplatform (FDEP NPs) by integration of doxorubicin (DOX) and epigallocatechin-3-O-gallate (EGCG) with the help of coordination between Fe 3+ ions and polyphenols. The EGCG from FDEP NPs could inhibit the expression of carbonyl reductase 1 (CBR1) protein and thereby inhibit the doxorubicinol (DOXOL) generation from DOX both in vitro and in vivo , thus the efficacy of DOX to cancerous cells is improved significantly. More importantly, the FDEP NPs could reduce cardiac toxicity and the DOX mediated toxicity to blood cells due to the repression of DOXOL production. Moreover, the blood half-life of FDEP NPs is longer than 23 h as determined by positron emission tomography (PET) imaging of biodistribution of radiolabelled NPs and HPLC measurement of plasma level of DOX, ensuring high tumor accumulation of FDEP NPs by enhanced permeability and retention (EPR) effect. The FDEP NPs also exhibited much improved antitumor effect over free drugs. Our work sheds new light on the engineering of nanomaterials for combination chemotherapy and may find unique clinical applications in the near future.

As highly expressed in insulinomas, the glucagon-like peptide-1 receptor (GLP-1R) is believed to be an attractive target for diagnosis, localization, and treatment with radiolabeled exendin 4. However, the high and persistent radioactivity accumulation of exendin 4 in the kidneys limits accurate diagnosis and safe, as well as effective, radiotherapy in insulinomas. In this study, we intend to reduce the renal accumulation of radiolabeled exendin 4 through degradation mediated by brush border membrane enzymes. A new exendin 4 ligand NOTA-MVK-Cys40-Leu14-Exendin 4 containing Met-Val-Lys (MVK) linker between the peptide and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) chelator was synthesized and labeled with 68Ga. The in vitro mouse serum stability and cell binding affinity of the tracer were evaluated. Initial in vitro cleavage of the linker was determined by incubation of a model compound Boc-MVK-Dde with brush border membrane vesicles (BBMVs) with and without the inhibitor of neutral endopeptidase (NEP). Further cleavage studies were performed with the full structure of NOTA-MVK-Cys40-Leu14-Exendin 4. Kidney and urine samples were collected in the in vivo metabolism study after intravenous injection of 68Ga-NOTA-MVK-Cys40-Leu14-Exendin 4. The microPET images were acquired in INS-1 tumor model at different time points; the radioactivity uptake of 68Ga-NOTA-MVK-Cys40-Leu14-Exendin 4 in tumor and kidneys were determined and compared with the control radiotracer without MVK linker. 68Ga-NOTA-MVK-Cys40-Leu14-Exendin 4 was stable in mouse serum. The MVK modification did not affect the affinity of NOTA-MVK-Cys40-Leu14-Exendin 4 toward GLP-1R. The in vitro cleavage study and in vivo metabolism study confirmed that the MVK sequence can be recognized by BBM enzymes and cleaved at the amide bond between Met and Val, thus releasing the small fragment containing Met. MicroPET images showed that the tumor uptake of 68Ga-NOTA-MVK-Cys40-Leu14-Exendin 4 was comparable to that of the control, while the kidney uptake was significantly reduced. As a result, more favorable tumor to kidney ratios were achieved. In this study, a novel exendin 4 analogue, NOTA-MVK-Cys40-Leu14-Exendin 4, was successfully synthesized and labeled with 68Ga. With the cleavable MVK sequence, this ligand could be cleaved by the enzymes on kidneys, and releasing the fragment of 68Ga-NOTA-Met-OH, which will rapidly excrete from urine. As the high and consistent renal radioactivity accumulation could be significantly reduced, NOTA-MVK-Cys40-Leu14-Exendin 4 shows great potential in the diagnosis and radiotherapy for insulinoma.

Magnetic resonance imaging (MRI) diagnosis is better assisted by contrast agents that can augment the signal contrast in the imaging appearance. However, this technique is still limited by the inherently low sensitivity on the recorded signal changes in conventional T1 or T2 MRI in a qualitative manner. Here, we provide a new paradigm of MRI diagnosis using T1- T2 dual-modal MRI contrast agents for contrast-enhanced postimaging computations on T1 and T2 relaxation changes. An albumin-binding molecule (i.e., truncated Evans blue) chelated with paramagnetic manganese ion was developed as a novel T1- T2 dual-modal MRI contrast agent at high magnetic field (7 T). Furthermore, the postimaging computations on T1- T2 dual-modal MRI led to greatly enhanced signal-to-noise ratios (SNR) and contrast-to-noise ratios (CNR) in both subcutaneous and orthotopic brain tumor models compared with traditional MRI methods. The T1- T2 dual-modal MRI computations have great potential to eliminate suspicious artifacts and false-positive signals in mouse brain imaging. This study may open new avenues for contrast-enhanced MRI diagnosis and holds great promise for precision medicine.

Tumour hypoxia, the “Achilles’ heel” of current cancer therapies, is indispensable to drug resistance and poor therapeutic outcomes especially for radiotherapy. Here we propose an in situ catalytic oxygenation strategy in tumour using porphyrinic metal-organic framework (MOF)-gold nanoparticles (AuNPs) nanohybrid as a therapeutic platform to achieve O(2)-evolving chemoradiotherapy. The AuNPs decorated on the surface of MOF effectively stabilize the nanocomposite and serve as radiosensitizers, whereas the MOF scaffold acts as a container to encapsulate chemotherapeutic drug doxorubicin. In vitro and in vivo studies verify that the catalase-like nanohybrid significantly enhances the radiotherapy effect, alleviating tumour hypoxia and achieving synergistic anticancer efficacy. This hybrid nanomaterial remarkably suppresses the tumour growth with minimized systemic toxicity, opening new horizons for the next generation of theranostic nanomedicines.

The success of radiotherapy relies on tumor-specific delivery of radiosensitizers to attenuate hypoxia resistance. Here we report an ammonia-assisted hot water etching strategy for the generic synthesis of a library of small-sized (sub-50 nm) hollow mesoporous organosilica nanoparticles (HMONs) with mono, double, triple, and even quadruple framework hybridization of diverse organic moieties by changing only the introduced bissilylated organosilica precursors. The biodegradable thioether-hybridized HMONs are chosen for efficient co-delivery of tert-butyl hydroperoxide (TBHP) and iron pentacarbonyl (Fe(CO)5). Distinct from conventional RT, radiodynamic therapy (RDT) is developed by taking advantage of X-ray-activated peroxy bond cleavage within TBHP to generate •OH, which can further attack Fe(CO)5 to release CO molecules for gas therapy. Detailed in vitro and in vivo studies reveal the X-ray-activated cascaded release of •OH and CO molecules from TBHP/Fe(CO)5 co-loaded PEGylated HMONs without reliance on oxygen, which brings about remarkable destructive effects against both normoxic and hypoxic cancers., A common failure of many cancer treatments is attributed to the resistance imparted by tumour hypoxia. Here, the authors report on the generic synthesis of small-sized hollow mesoporous organosilica nanoparticles which are designed for oxygen-independent X-ray-activated synergistic therapy.

It remains a major challenge to achieve precise on-demand drug release. Here, we developed a modular nanomedicine integrated with logic-gated system enabling programmable drug release for on-demand chemotherapy. Methods: We employed two different logical AND gates consisting of four interrelated moieties to construct the nanovesicles, denoted as v-A-CED2, containing oxidation-responsive nanovesicles (v), radical generators (A), and Edman linker conjugated prodrugs (CED2). The first AND logic gate is connected in parallel by mild hyperthermia ( I ) and acidic pH ( II ), which executes NIR laser triggered prodrug-to-drug transformation through Edman degradation. Meanwhile, the mild hyperthermia effect triggers alkyl radical generation ( III ) which contributes to internal oxidation and degradation of nanovesicles ( IV ). The second AND logic gate is therefore formed by the combination of I-IV to achieve programmable drug release by a single stimulus input NIR laser. The biodistribution of the nanovesicles was monitored by positron emission tomography (PET), photoacoustic, and fluorescence imaging. Results: The developed modular nanovesicles exhibited high tumor accumulation and effective anticancer effects both in vitro and in vivo. Conclusions: This study provides a novel paradigm of logic-gated programmable drug release system by a modular nanovesicle, which may shed light on innovation of anticancer agents and strategies.

The design of hybrid metal-organic framework (MOF) nanomaterials by integrating inorganic nanoparticle into MOF (NP@MOF) has demonstrated outstanding potential for obtaining enhanced, collective, and extended novel physiochemical properties. However, the reverse structure of MOF-integrated inorganic nanoparticle (MOF@NP) with multifunction has rarely been reported. Methods: We developed a facile in-situ growth method to integrate MOF nanoparticle into inorganic nanomaterial and designed a fluorescence switch to trigger enhanced photodynamic therapy. The influence of “switch” on the photodynamic activity was studied in vitro. The in vivo mice with tumor model was applied to evaluate the “switch”-triggered enhanced photodynamic therapy efficacy. Results: A core-satellites structure with fluorescence off and on function was obtained when growing MnO2 on the surface of fluorescent zeolitic imidazolate framework (ZIF-8) nanoparticles. Furthermore, A core-shell structure with photodynamic activity off and on function was achieved by growing MnO2 on the surface of porphyrinic ZrMOF nanoparticles (ZrMOF@MnO2). Both the fluorescence and photodynamic activities can be turned off by MnO2 and turned on by GSH. The GSH-responsive activation of photodynamic activity of ZrMOF@MnO2 significantly depleted the intracellular GSH via a MnO2 reduction reaction, thus triggering an enhanced photodynamic therapy efficacy. Finally, the GSH-reduced Mn2+ provided a platform for magnetic resonance imaging-guided tumor therapy. Conclusion: This work highlights the impact of inorganic nanomaterial on the MOF properties and provides insight to the rational design of multifunctional MOF-inorganic nanomaterial complexes.

Biomedical applications and nanotechnological advances, including constrained synthesis, multimodal imaging, drug delivery, and bioassay, have strongly benefited from employing ferritin nanocages due to their remarkable properties of easy engineering, great biocompatible features, large capacity and so on. In this study, ferritin nanocages were used to display epitopes (model antigens derived from Enterovirus 71 (EV71) with different length) on C- and N-terminals and the loop zone to search for the optimal position for the fusion of the epitopes to the vaccine platform. The longest epitope displayed on the N-terminal and loop zone as well as the second longest peptide displayed on the loop zone of ferritin resulted in 100% passive protection of newborn BALB/c mice from the lethal EV71. This suggests that peptides fused onto the loop zone of ferritin could induce strong immune response. Our results increase the versatility of the vaccine platform and provide more options for the production of stable constructs, suggesting the potential future clinical applicability of ferritin-based antigen delivery nanoplatforms.

A Pt prodrug polyphenol and gadolinium ion loaded cancer theranostics nanoplatform based on mild acidic pH and thermal sensitive polymer was designed for photoacoustic (PA)/ magnetic resonance(MR)/ positron emission tomography (PET) multimodal imaging-guided chemo-photothermal combination therapy. The Pt drug release can be controlled by tumour-specific acidic pH and heat generated by external NIR irradiation. The nanoparticles were stable under normal physiological environment and released the drug under tumour acidic pH and NIR laser irradiation, which can reduce the side effect of drug to normal organs. Moreover, the MR signal can be significantly enhanced (~3-fold increase in T1 relaxivity) under the acidic tumour microenvironment, which is favorable for cancer diagnosis. The nanoparticles exhibited excellent tumour accumulation and led to complete tumour eradication with low power NIR laser irradiation. This promising approach provides a new avenue for imaging-guided combination therapy.

Nanomedicines with photodynamic therapy and reactive oxygen species (ROS)‐triggered drug release capabilities are promising for cancer therapy. However, most of the nanomedicines based on ROS‐responsive nanocarriers still suffer from serious ROS consumption during the triggered drug release process. Herein, a photodynamic‐chemodynamic cascade strategy for the design of drug delivery nanosystem is proposed. A doxorubicin hydrochloride‐loaded ROS‐responsive polymersome (DOX‐RPS) is prepared via the self‐assembly of amphiphilic poly(ethylene glycol)‐poly(linoleic acid) and poly(ethylene glycol)‐(2‐(1‐hexyloxyethyl)‐2‐devinyl pyropheophorbide‐α)‐iron chelate (PEG‐HPPH‐Fe). The RPS can effectively deliver a drug to tumor site through passive targeting effect. Upon laser irradiation, the photosensitizer HPPH can efficiently generate ROS, which further causes in situ oxidation of linoleic acid chain and subsequent RPS structural destruction, permitting triggered drug release. Intriguingly, catalyzed by HPPH‐Fe, ROS will be regenerated from linoleic acid peroxide through a chemodynamic process. Therefore, ROS‐triggered drug release can be achieved without ROS over‐consumption. The in vitro and in vivo results confirmed ROS generation, triggered drug release behavior, and potent antitumor effect of the DOX‐RPS. This photodynamic‐chemodynamic cascade strategy provides a promising approach for enhanced combination therapy., Photodynamic‐chemodynamic cascade strategy: A reactive oxygen species (ROS)‐responsive polymersome is developed for chemo‐photodynamic combination cancer therapy. The ROS generated by photodynamic effect will cause ROS‐triggered drug release. Furthermore, ROS could be regenerated through a Fenton‐like reaction. Therefore, excessive ROS consumption in the drug release process can be avoided.

The outcome of radiotherapy is significantly restricted by tumor hypoxia. To overcome this obstacle, one prevalent solution is to increase intratumoral oxygen supply. However, its effectiveness is often limited by the high metabolic demand for O2 by cancer cells. Herein, we develop a hybrid semiconducting organosilica-based O2 nanoeconomizer pHPFON-NO/O2 to combat tumor hypoxia. Our solution is twofold: first, the pHPFON-NO/O2 interacts with the acidic tumor microenvironment to release NO for endogenous O2 conservation; second, it releases O2 in response to mild photothermal effect to enable exogenous O2 infusion. Additionally, the photothermal effect can be increased to eradicate tumor residues with radioresistant properties due to other factors. This “reducing expenditure of O2 and broadening sources” strategy significantly alleviates tumor hypoxia in multiple ways, greatly enhances the efficacy of radiotherapy both in vitro and in vivo, and demonstrates the synergy between on-demand temperature-controlled photothermal and oxygen-elevated radiotherapy for complete tumor response., Tumor hypoxia is a major limitation in radiotherapy, and strategies to address this often fail due to high oxygen consumption. Here, the authors report a nanomaterial assembly for the simultaneous reduction in mitochondrial respiration and to supply oxygen to potentiate radiotherapy.

The combination of reactive oxygen species (ROS)-involved photodynamic therapy (PDT) and chemodynamic therapy (CDT) holds great promise for enhancing ROS-mediated cancer treatment. Herein, we reported an in situ polymerized hollow mesoporous organosilica nanoparticle (HMON) biocatalysis nanoreactor to integrate the synergistic effect of PDT/CDT for enhancing ROS-mediated pancreatic ductal adenocarcinoma treatment. HPPH photosensitizer was hybridized within the framework of HMON via an “in situ framework growth” approach. Then, the hollow cavity of HMONs was exploited as a nanoreactor for “in situ polymerization” to synthesize the polymer containing thiol groups, thereby enabling the immobilization of ultrasmall gold nanoparticles, which behave like glucose oxidase-like nanozyme, converting glucose into H(2)O(2) to provide self-supplied H(2)O(2) for CDT. Meanwhile, Cu(2+)-tannic acid complexes were further deposited on the surface of HMONs (HMON-Au@Cu-TA) to initiate Fenton-like reaction to covert the self-supplied H(2)O(2) into •OH, a highly toxic ROS. Finally, collagenase (Col), which can degrade the collagen I fiber in the extracellular matrix (ECM), was loaded into HMON-Au@Cu-TA to enhance the penetration of HMONs and O(2) infiltration for enhanced PDT. This study provides a good paradigm for enhancing ROS-mediated anti-tumor efficacy. Meanwhile, this research offers a new method to broaden the application of silica based nanotheranostics.

Tumor heterogeneity is one major reason for unpredictable therapeutic outcomes, while stratifying therapeutic responses at an early time may greatly benefit the better control of cancer. Here, we developed a hybrid nanovesicle to stratify radiotherapy response by activatable inflammation magnetic resonance imaging (aiMRI) approach. The high Pearson’s correlation coefficient R values are obtained from the correlations between the T1 relaxation time changes at 24–48 h and the ensuing adaptive immunity (R = 0.9831) at day 5 and the tumor inhibition ratios (R = 0.9308) at day 18 after different treatments, respectively. These results underscore the role of acute inflammatory oxidative response in bridging the innate and adaptive immunity in tumor radiotherapy. Furthermore, the aiMRI approach provides a non-invasive imaging strategy for early prediction of the therapeutic outcomes in cancer radiotherapy, which may contribute to the future of precision medicine in terms of prognostic stratification and therapeutic planning., Non-invasive approaches to stratify early responses to therapy are of great value to improve cancer patient management. Here the authors design a hybrid nanovesicle-based activatable inflammation magnetic resonance method for the early prediction of response to radiotherapy.

Background: Radiolabeled somatostatin (SST) analogs have been proven to be effective in the diagnosis and treatment of neuroendocrine tumors (NETs), which are characterized by high somatostatin receptor (SSTR2) expression. At present, there are several SST analogs available that differ from each other in the affinity for SSTR2 and tumor retention time. To date, only a single SST agonist -DOTA-TATE- has been approved by the FDA for imaging and treatment of NETs. Recent studies have shown that addition of Evans blue (EB) moiety to an SST agonist results in superior uptake and increased retention time within the tumors. The goal of our study was to compare the diagnostic and therapeutic efficacy of three different radiolabeled SST analogs in a tumor mice model: EB-TATE - a novel modified agonist; DOTA-TATE - an agonist and JR11 - an antagonist. Methods: A rat pancreatic cell line (AR42J), characterized by high SSTR2 expression, was used to create a subcutaneous xenograft mice model. The AR42J cells formed sizable tumors within two weeks post-injection. The 86Y-EB-TATE, 68Ga-DOTA-TATE, and 68Ga-DOTA-JR11 were used to determine standard uptake values by positron emission tomography (PET) imaging. For treatment purposes, the SST analogs were labeled with 177Lu to generate 177Lu-EB-TATE, 177Lu-DOTA-TATE and 177Lu-DOTA-JR11. The mice were assigned to treatment groups based on comparable tumor volume at baseline and received two doses (0.5mCi) of the 177Lu-labeled analogs one week apart. Tumor measurements were performed twice per week and the mice were euthanized if their tumor burden exceeded 2 cm at any point in the study or after 6 weeks - landmark of the end of the study. Results: Among the three analogs tested, the novel SST analog 86Y-EB-TATE was characterized by 4.3- and 3.7- fold higher tumor uptake in comparison to 68Ga-DOTA-TATE (p