ImmunoPET imaging of Trop2 expression in solid tumors with nanobody tracers.

PURPOSE: The high expression of the transmembrane glycoprotein trophoblast cell-surface antigen 2 (Trop2) was strongly associated with the progression of solid tumors, including pancreatic and gastric cancers. Our study aimed to construct Trop2-specific immuno-positron emission tomography (immunoPET) probes and assess the diagnostic abilities in preclinical pancreatic and gastric cancer models. METHODS: The expression of Trop2 in pancreatic cancer was determined by single-cell sequencing and immunohistochemistry on tissue microarray (TMA). Flow cytometry was used to screen the expression of Trop2 in pancreatic cancer cell lines. Two nanobodies (i.e., RTD98 and RTD01) targeting Trop2 were developed and labeled with gallium-68 (68Ga, T1/2 = 1.1 h) to construct immunoPET imaging probes. The agents were researched in cell-derived pancreatic and patient-derived gastric cancer models expressing varying Trop2. RESULTS: Single-cell sequencing results showed high expression of Trop2 in pancreatic ductal cells as well as acinar cells and immunohistochemical staining of TMA from pancreatic cancers showed significantly higher expression of Trop2 in cancerous than in paracancerous tissues. ImmunoPET utilizing [68Ga]Ga-NOTA-RTD98 could clearly delineate subcutaneous tumors, both in cell-derived pancreatic cancer models and patient-derived gastric cancer models, superior to imaging using [18F]-FDG or a non-specific probe [68Ga]Ga-NOTA-RTD161. Another probe with improved pharmacokinetics targeting Trop2, [68Ga]Ga-NOTA-RTD01, was further prepared and showed advantageous diagnostic capabilities in preclinical pancreatic cancer models. CONCLUSION: In the work, we reported two nanobody tracers targeting human Trop2 which may facilitate better use of Trop2-targeted therapeutics by noninvasively displaying expression dynamics of the target.

Interfacial Self-Assembly of Surfactant-Free Au Nanoparticles as a Clean Surface-Enhanced Raman Scattering Substrate for Quantitative Detection of As5+ in Combination with Convolutional Neural Networks.

Surface-enhanced Raman scattering (SERS) is a highly sensitive tool in the field of environmental testing. However, the detection and accurate quantification of weakly adsorbed molecules (such as heavy metal ions) remain a challenge. Herein, we combine clean SERS substrates capable of capturing heavy metal ions with convolutional neural network (CNN) algorithm models for quantitative detection of heavy metal ions in solution. The SERS substrate consists of surfactant-free Au nanoparticles (NPs) and l-cysteine molecules. As plasmonic nanobuilt blocks, surfactant-free Au NPs without physical or chemical barriers are more accessible to target molecules. The amino and carboxyl groups in the l-cysteine molecule can chelate As5+ ions. The CNN algorithm model is applied to quantify and predict the concentration of As5+ ions in samples. The results demonstrated that this strategy allows for fast and accurate prediction of As5+ ion concentrations, and the determination coefficient between the predicted and actual values is as high as 0.991.

Superior biocompatible carbon dots for dynamic fluorescence imaging of nucleoli in living cells.

The nucleolus is a newly developed and promising target for cancer diagnosis and therapy, and its imaging is extremely significant for fundamental research and clinical applications. The unique feature, i.e., high resolution at the subcellular level, makes the fluorescence imaging method a powerful tool for nucleolus imaging. However, the fluorescence imaging of nucleoli in living cells is restricted by the limited availability of fluorescent agents with specific nucleolus-targeting capability and superior biocompatibility. Here, promising carbon dots (CDs) with intrinsic nucleolus-targeting capability were synthesized, characterized and employed for dynamic fluorescence imaging of nucleoli in living cells. The CDs exhibit a high fluorescence quantum yield of 0.2, excellent specificity and photostability, and superior biocompatibility, which were systematically demonstrated at the gene, cellular and animal levels and confirmed by their biological effects on embryonic development. All these features enabled CDs to light up the nucleoli for a long time with a high signal-to-noise ratio in living cells and monitor the nucleolar dynamics of malignant cells in camptothecin (CPT) based chemotherapy. Their excellent optical and biological features as well as general nucleolus-targeting capability endow CDs with great potential for future translational research.

Hyperandrogenism caused by a rare adrenocortical oncocytic neoplasm with uncertain malignant potential: a case report and review of the literature.

Hyperandrogenism is a state of androgen excess that can induce hirsutism and oligo/amenorrhea in women of reproductive age. Therapeutic strategies differ according to etiology. Hence, the differential diagnosis of hyperandrogenism is crucial. The adrenal gland is an important organ that produces androgens. One common cause of hyperandrogenism is androgen-secreting adrenal tumors; however, adrenocortical oncocytic neoplasms (ACONs) are rare. A 23-year-old woman presented with severe hirsutism and menstrual disorders for 2 years. Her Ferriman-Gallway hirsutism score was 15 at her first consultation. Her menstrual cycles were irregular, and her menstrual flow had diminished gradually over the past 2 years. She had a remarkable elevation of total testosterone, dehydroepiandrosterone sulfate and androstenedione. Pelvic ultrasonography showed normal morphology of the uterus and bilateral ovaries. Computed tomography revealed a giant left adrenal tumor with a diameter of 12 cm. The patient then underwent robotic-assisted adrenal tumor resection. Histopathological assessment indicated adrenocortical oncocytic neoplasm with uncertain malignant potential. After 4 years of follow-up, no recurrence of symptoms was noted, and this patient delivered a healthy infant on her due date in October 2021. This article reviews the clinical features, diagnosis, and treatment of ACONs and highlights the importance of differential diagnosis for hyperandrogenism in women.

Ratiometric Raman nanotags enable intraoperative detection of metastatic sentinel lymph node.

Sentinel lymph node (SLN) imaging and biopsy has been advocated as an important technique to evaluate the metastatic status of regional lymph nodes and determine subsequent surgical procedure for many cancers, yet there is no reliable means to provide accurate and rapid diagnosis of metastatic SLN during surgery. Here we develop a new approach, named "Ratiometric Raman dual-nanotag strategy", that using folic acid functionalized targeted and nontargeted gap-enhanced Raman tags (FA-GERTs and Nt-GERTs) to detect metastatic SLN based on Raman imaging combined with classical least square data processing methods. By using this strategy, with built-in self-calibration for signal correction, rather than absolute intensity-dependent signal readout, we realize the visualization and prompt intraoperative diagnosis of metastatic SLN with a high accuracy of 87.5 % when the cut-off value of ratio (FA-GERTs/Nt-GERTs) set at 1.255. This approach may outperform the existing histopathological assessment in diagnosing SLN metastasis and is promising for guiding surgical procedure in the future.

Nucleolus-Targeted Photodynamic Anticancer Therapy Using Renal-Clearable Carbon Dots.

Photodynamic therapy (PDT), which utilizes light excited photosensitizers (PSs) to generate reactive oxygen species (ROS) and consequently ablate cancer cells or diseased tissue, has attracted a great deal of attention in the last decades due to its unique advantages. In order to further enhance PDT effect, PSs are functionalized to target specific sub-cellular organelles, but most PSs cannot target nucleolus, which is demonstrated as a more efficient and ideal site for cancer treatment. Here, an effective carbon dots (C-dots) photosensitizer with intrinsic nucleolus-targeting capability, for the first time, is synthesized, characterized, and employed for in vitro and in vivo image-guided photodynamic anticancer therapy with enhanced treatment performance at a low dose of PS and light irradiation. The C-dots possess high ROS generation efficiency and fluorescence quantum yield, excellent in vitro and in vivo biocompatibility, and rapid renal clearance, endowing it with a great potential for future translational research.

Intraoperative Raman-Guided Chemo-Photothermal Synergistic Therapy of Advanced Disseminated Ovarian Cancers.

Abdominal miliary spread and metastasis is one of the most aggressive features in advanced ovarian cancer patients. The current standard treatment of advanced ovarian cancer is cytoreductive surgery (CRS) combined with hyperthermic intraperitoneal chemotherapy (HIPEC). However, most patients cannot receive optimal CRS outcomes due to the extreme difficulty of completely excising all microtumors during operation. Though HIPEC can improve prognosis, treatment is untargeted and may damage healthy organs and cause complications. New strategies for precise detection and complete elimination of disseminated microtumors without side effects are therefore highly desirable. Here, cisplatin-loaded gap-enhanced Raman tags (C-GERTs) are designed specifically for the intraoperative detection and elimination of unresectable disseminated advanced ovarian tumors. With unique and strong Raman signals, good biocompatibility, decent plasmonic photothermal conversion, and good drug loading capacity, C-GERTs enable detection and specific elimination of microtumors with a minimum diameter of 1 mm via chemo-photothermal synergistic therapy, causing minimal side effects and significantly prolonging survival in mice. The results demonstrate that C-GERTs-based chemo-photothermal synergistic therapy can effectively control the spread of disseminated tumors in mice and has potential as a safe and powerful method for treatment of advanced ovarian cancers, to improve survival and life quality of patients.

Gap-enhanced Raman tags for high-contrast sentinel lymph node imaging.

The sentinel lymph node (SLN) biopsy is gaining in popularity as a procedure to investigate the lymphatic metastasis of malignant tumors. The commonly used techniques to identify the SLNs in clinical practice are blue dyes-guided visualization, radioisotope-based detection and near-infrared fluorescence imaging. However, all these methods have not been found to perfectly fit the clinical criteria with issues such as short retention time in SLN, poor spatial resolution, autofluorescence, low photostability and high cost. In this study, we have reported a new type of nanoprobes, named, gap-enhanced Raman tags (GERTs) for the SLN Raman imaging. With the advantageous features including unique "fingerprint" Raman signal, strong Raman enhancement, high photostability, good biocompatibility and extra-long retention time, we have demonstrated that GERTs are greatly favorable for high-contrast and deep SLN Raman imaging, which meanwhile reveals the dynamic migration behavior of the probes entering the SLN. In addition, a quantitative volumetric Raman imaging (qVRI) data-processing method is employed to acquire a high-resolution 3-dimensional (3D) margin of SLN as well as the content variation of GERTs in the SLN. Moreover, SLN detection could be realized via a cost-effective commercial portable Raman scanner. Therefore, GERTs hold the great potential to be translated in clinical application for accurate and intraoperative location of the SLN.