Figure S8. Quantification of manganese content in the PANC1 tumor 60 minutes post-injection of MnL3. MnL3 exhibited significantly higher tumor uptake in PANC1 tumors treated with one dose of FOLFIRINOX compared to the untreated mice (0.1 mmol/kg, i.v., 60 min p.i., n = 4 for each group, Student’s t-test, two-tailed, *P<0.05).

Early, noninvasive, and spatially resolved assessment of renal dysfunction is essential for theranostics of kidney diseases, yet clinical biomarkers are criticized for averaging whole-kidney function and missing focal or unilateral injury. Here, we present a contrast-sparing computed tomography (CT) approach that maps renal dysfunction at the compartment level using renal-clearable glutathione-gold nanoclusters (GSH-Au NCs). The merits of ultrasmall hydrodynamic size (2.8 nanometers) and high x-ray attenuation ability enable GSH-Au NCs with fast renal clearance and robust intrarenal CT contrast. In three mechanistically distinct mouse models of acute kidney injury, low-dose GSH-Au NCs produced mechanism-specific imaging signatures and distinct time-attenuation behaviors, even before biomarkers changed. Notably, at an ultralow dose (32.5 milligrams of Au per kilogram), early bilateral cisplatin injury can be easily detected with an obvious bright band at the outer medulla area on CT, consistent with tubular injury and a cast/debris-mediated mechanism. This work establishes a dose-efficient CT workflow for early, noninvasive, spatial mapping of renal dysfunction, supporting diagnosis, classification, and monitoring of kidney disease.

Figure S10. Quantification of manganese content in the 1319 tumor 60 minutes post-injection of MnL3. After randomly assigning 1319 mice to the vehicle or FOLFIRINOX group, MnL3 showed equivalent tumor uptake (0.1 mmol/kg, i.v., 60 min p.i., n = 5 for each group, Student’s t-test. ns: not significant).

Abstract Fibrogenesis is essential to wound healing, but aberrant fibrogenesis is a driver of many chronic diseases and cancers. Lysyl oxidases (LOX) play a pivotal role in fibrogenesis by catalyzing the oxidation of lysine residues to reactive aldehydes (allysine) in collagens and elastin, resulting in the crosslinking and excessive deposition of these extracellular matrix components. Currently, rapid and robust histological assays to visualize the spatial distribution of LOX activity are lacking, hindering the precise validation of anti-fibrotic therapies. Here, we present a histological fluorescent staining method to visualize fibrogenesis (active fibrosis) and LOX activity in tissue sections utilizing a bioorthogonal tag and a click reaction with a turn-on fluorophore. Notably, requiring only two commercial reagents, this protocol can be completed in under two hours and is compatible with other imaging modalities, including second-harmonic generation and immunofluorescence staining. We validated this method across various healthy and fibrotic mouse and human tissue specimens.

Figure S5. Immediate dynamic contrast enhanced (DCE) MRI in PDAC6 tumor with MnL4 performed before, at 3 days (1 dose FOLFIRNOX), or at 7 days (2 doses FOLFIRINOX) after the initiation of FOLFIRINOX treatment. Time course of change in normalized signal intensity acquired from DCE images with MnL4 as the probe (0.1 mmol/kg, i.v., n = 4 for each group, data shown as mean value with 95% simultaneous confidence bands as shaded regions). Data are superimposable at each time point measured, indicating no changes in tumor permeability or extracellular volume with FOLFIRINOX treatment.

Figure S4. The PDAC6 tumor percentage change of normalized signal intensity (%nSI, normalized to an adjacent phantom) at 30 min p.i. of probe (n = 4 for each group). One-way ANOVA with Tukey’s post hoc test, *P<0.05, **P < 0.01, ns, not significant.

The successful management of type-1 diabetes (T1D) and insulin-dependent type-2 diabetes depends on the ability to accurately calibrate bolus and basal insulin doses, and minimize the time spent with high postprandial blood glucose (BG) levels while avoiding dangerously low hypoglycemic excursions. Precise calibration of the insulin pump helps with successful disease management; however, people with T1D may still experience prolonged, potentially damaging BG levels due to postprandial highs. Insulin pumps offer large, currently underexploited degrees of freedom in insulin delivery, which can have a dramatic impact on average postprandial BG levels. Building on existing models of the glucose-insulin system, we first propose a simple, automated, and individualized insulin-pump calibration system based on a series of measures taken before and a few hours after carbohydrate ingestion. We then modulate the shape of pre-meal insulin dosing to explicitly reduce postprandial BG levels while minimizing the likelihood of dangerously low BG. Such an optimal insulin delivery time course can potentially improve postprandial BG levels and rapidly bring BG to the target level. We evaluated our methods in 20 patients over 4 days, with BG levels sampled frequently.

Figure S7. MnL3 molecular MRI of PANC1 tumor-bearing mice before administration of vehicle or FOLFIRINOX. After randomly assigning PANC1 mice to FOLFIRINOX or vehicle group, MnL3 molecular MRI (0.1 mmol/kg, i.v., 60 min p.i.) showed equivalent tumor enhancement before administration of vehicle or FOLFIRINOX. (n = 8 for each group, Student’s t-test, two-tailed, ns, not significant.).

Figure S2. Mass spectra of MnL4 before and after incubation with butyraldehyde. (A) MS spectrum of MnL4. [M + H]+ calcd for [C25H39MnN6O7], 590.2; found 590.0. (B) MnL4 did not react with butyraldehyde at pH 7.4 (PBS). [M + H]+ calcd for [C25H39MnN6O7], 590.2; found 590.0). (C) MnL4 did not react with butyraldehyde at pH 6.5 (PBS). [M + H]+ calcd for [C25H39MnN6O7], 590.2; found 590.0).

Macrophage density and phenotype are key components of the innate immune response and have been implicated in the outcomes and progression of cancer and autoimmune diseases. The ability to quantify and perform temporospatial analysis of macrophages could enable more specific therapeutic interventions. Macrin, a 17-nm nanoparticle, has shown high selectivity for macrophage accumulation in preclinical models of disease. The purpose of this first-in-human study was to evaluate the biodistribution, dosimetry, and safety of 64Cu-labeled Macrin and to explore its potential for imaging inflammatory and malignant conditions. Methods: This study (NCT04843891) enrolled 10 human subjects (7 healthy volunteers and 3 patient subjects) who received a single administration of 64Cu-Macrin. The mean administered activity was approximately 444 MBq. Serial PET/CT scans were performed to determine the whole-body biodistribution and to calculate radiation dosimetry. Blood samples were collected at multiple time points to assess the pharmacokinetic profile. Participants were monitored for any adverse events or pharmacologic effects. Results: No adverse or clinically detectable pharmacologic effects were observed in any of the 10 participants, establishing the safety of the imaging agent. The blood clearance of 64Cu-Macrin was biphasic, with a fast component half-life of 1.3 h and a slow component of 22.3 h. In participants with diagnosed cancer and sarcoidosis, notable accumulation of 64Cu-Macrin was observed at sites of active disease. Conclusion: The results of this first-in-human study support the safety and favorable pharmacokinetic profile of 64Cu-Macrin PET imaging. The tracer successfully accumulated in macrophage-rich tissues, such as sites of cancer and sarcoidosis, suggesting that this approach could be a valuable tool for patient stratification and therapy response assessment, particularly in the development of macrophage-targeted therapeutics.

To evaluate the feasibility of collagen-binding ⁶⁸Ga-CBP8 PET/MR for the noninvasive detection of deep infiltrating endometriosis. A single-case exploratory application of ⁶⁸Ga-CBP8 PET/MR was conducted in a patient with endometriosis as a preliminary step toward a larger IRB-approved prospective study. Imaging was performed 60 min after intravenous administration of ⁶⁸Ga-CBP8. Fused PET/MR images were qualitatively assessed for focal tracer uptake exceeding background. Surgical pathology, assessed in a blinded fashion, served as the reference standard. A 32-year-old woman with history of endometriosis completed the examination without adverse events. PET/MR demonstrated focal ⁶⁸Ga-CBP8 uptake corresponding on MR to a T2-hypointense plaque located between the left adnexa and the sigmoid colon, suspicious for deep pelvic endometriosis with bowel involvement. Surgical pathology confirmed endometriosis infiltrating the colonic muscularis mucosae and pericolonic tissue with dense fibrosis. ⁶⁸Ga-CBP8 PET/MR is a feasible and promising technique to noninvasively detect deep infiltrating endometriosis, warranting validation in larger prospective studies. ClinicalTrials.gov, NCT06377553. Date of first enrollment: February 5, 2025. Registered April 22, 2024.

Figure S1. Mass spectra of MnL3 solutions before and after reaction with butyraldehyde. (A) MS spectrum of MnL3. [M + H]+ calcd for [C20H32MnN7O5], 505.2; found 505.0. (B) MS spectrum of the reaction product between MnL3 and butyraldehyde at pH 7.4 (PBS). [M + H]+ calcd for [C24H38MnN7O5], 559.2; found 559.5. (C) MS spectrum of the reaction product between MnL3 and butyraldehyde at pH 6.5(PBS).

OBJECTIVES:To explore magnetic resonance imaging (MRI) and gallium-68 ( 68 Ga)-DOTATATE positron emission tomography (PET) performance in the assessment of neuroendocrine liver metastases (NELMs) on a per-lesion basis, with particular attention to the contribution of individual MRI sequences and assessment of other factors that might influence their detection. MATERIALS AND METHODS:This observational retrospective study included patients with histologically confirmed neuroendocrine tumors who underwent both contrast-enhanced MRI and 68 Ga-DOTATATE PET within 12 weeks between August 2017 and December 2023. Three readers in consensus assessed individual MRI sequences [diffusion-weighted imaging (DWI), dynamic contrast-enhanced imaging (DCE), and hepatobiliary phase (HBP) imaging when available], entire MRI data set, and PET in random order. The reference standard was histopathology or follow-up imaging. Diagnostic performance metrics were calculated using generalized estimating equations with Bonferroni correction. Correlations were assessed using Pearson correlation coefficients. RESULTS:A total of 1249 lesions, comprising 1050 metastases, were analyzed in 60 patients (mean age: 64.9±11.5 years; 56.7% male). Compared with PET, MRI demonstrated superior sensitivity (93% vs. 59%, P <0.001) and accuracy (93% vs. 63%, P <0.001), with DWI and HBP providing the highest sensitivity (89% and 92%). Size-stratified analysis showed that MRI outperformed PET, particularly for metastases <5 mm (81.6% vs. 19.7%) and 5 to 10 mm (96.1% vs. 61.8%) ( P <0.001). Arterial enhancement and portal venous washout were present in 67.8% and 23.7% of metastases, respectively, with only portal venous washout showing size dependence (11.9% in <5 mm to 55.6% in >20 mm lesions, P <0.01). PET-negative metastases were smaller than PET-positives (5.0 vs. 8.0 mm, P =0.001), with lesion size correlating with maximum standardized uptake values and normalized uptake ratios ( r =0.54 to 0.59, P <0.001). CONCLUSIONS:MRI outperformed 68 Ga-DOTATATE PET in detecting NELMs, with DWI and HBP providing particularly high sensitivity for small metastases.

Figure S6. Quantification of manganese content in the PDAC6 tumor 60 minutes post-injection of MnL3. MnL3 (0.1 mmol/kg, i.v., 60 min p.i.) exhibited significantly higher tumor uptake at 3 days (P = 0.05) and 7 days (P = 0.04) after initiation of FOLFIRINOX treatment compared to the untreated tumor (One-way ANOVA with Tukey’s post hoc test, *P<0.05, ns, not significant. n = 6 for day 0 and day 3, n = 5 for day 7).

Figure S9. MnL3 molecular MRI of 1319 tumor-bearing mice before administration of vehicle or FOLFIRINOX. After randomly assigning 1319 mice to the vehicle or FOLFIRINOX group, MnL3 molecular MRI showed equivalent tumor enhancement before administration of vehicle or FOLFIRINOX. ( n = 4 for each group, Student’s t-test. ns: not significant).

OBJECTIVES:Development of molecular therapies for liver fibrosis is slowed by a lack of noninvasive methods addressing questions of target expression, target engagement, and treatment response. Integrin α v β 6 is a biomarker of liver fibrosis that is upregulated in livers of patients with primary sclerosing cholangitis. It activates latent TGF-β and plays a critical role in regulating extracellular matrix expression, especially collagen. In this study, our aim was to use combined α v β 6 integrin-targeted positron emission tomography (PET) and collagen-specific magnetic resonance imaging (MRI) to measure target expression/engagement and liver fibrosis reduction with a α v β 6 integrin inhibitor. MATERIALS AND METHODS:We conducted a treatment study in bile duct-ligated (BDL) rats using a small molecule inhibitor to α v β 6 /α v β 1 . 68 Ga-DOTA-R01-MG, an α v β 6 -specific PET probe, was used to noninvasively measure α v β 6 expression and target engagement in the liver. CM-101, a type I collagen MRI probe, was used to quantify fibrosis. RESULTS:68 Ga-DOTA-R01-MG PET showed 3-fold higher liver uptake in BDL rats compared with sham rats at 17 days after surgery. Pretreatment with high dose α v β 6 /α v β 1 inhibitor 1 hour before imaging significantly decreased liver PET uptake in BDL rats (31%, P  = 0.012). Two weeks of daily dosing with an α v β 6 /α v β 1 inhibitor attenuated α v β 6 expression in BDL rat liver as assessed by α v β 6 PET (0.27 ± 0.07 percent injected dose [%ID]/mL compared with 0.40 ± 0.09 %ID/mL in vehicle-treated group, P  = 0.014) and reduced liver fibrosis as assessed by collagen MRI (liver relaxation rate change ΔR 1  = 0.14 ± 0.11 vs 0.36 ± 0.06, P  = 0.0037). Imaging findings were confirmed by histology (collagen proportionate area 10.7 ± 2.8% vs 22.5 ± 6.1%, P  < 0.001). CONCLUSIONS:A single imaging protocol combining molecular MRI and PET can be used to effectively monitor integrin inhibitor treatment by measuring target expression/engagement and treatment outcomes. Multimodality molecular imaging may be valuable in accelerating drug development in molecular therapies for liver fibrosis.

Detecting chronic lung allograft dysfunction (CLAD) remains difficult due to the lack of sensitive diagnostic markers. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) can quantify microvascular perfusion and extravascular extracellular space. We performed DCE-MRI on 10 participants with CLAD (4 BOS, 3 RAS, 1 mixed, 2 undefined) and 12 without CLAD, and calculated peak enhancement (PE), wash-in rate (kwashin), area under the first 60 s of the dynamic curve (iAUC60), and wash-out rate (kwashout) from signal intensity/time curves of lung regions of interest. Compared with participants without CLAD (age 61 [44-71]), those with CLAD (age 61 [54-68]) demonstrated a lower PE (p = 0.02), slower kwashin (p = 0.01), and reduced iAUC60 (p = 0.01), consistent with microvascular perfusion abnormalities. The kwashout was reduced in the lungs of CLAD participants, reflecting an increased extracellular extravascular space (p = 0.02). These findings support DCE-MRI as an important tool for detecting CLAD pathophysiology.