By: Geoffrey Feld, Ph.D. 
Geocyte LLC & Alamar Biosciences collaborator 

“This is personal to me. The early detection of cancer is the way to go.” 

In the July 2024 edition of the Mendelspod, Alamar Biosciences Founder Dr. Yuling Luo told Theral Timpson about how cancer took his mother and many other members of his family. Thus, Alamar was born with the goal of improving protein detection to enable early diagnosis of disease, particularly cancers.  

The promise of precision oncology 

Liquid biopsy is best known to tell whether you likely have cancer. Proteomics can potentially provide additional biomarker signatures of cancer detection, as well as clinically meaningful and personalized information, including progression monitoring and treatment response assessment (Pando-Caciano et al., 2024). 

But what if a routine fluid biosample (e.g., blood or urine) could also inform your: 

• Risk of developing organ toxicity upon receiving cancer immunotherapy  

• Likelihood of developing severe immune reactions upon CAR-T treatment 

• Risk of survival after cancer diagnosis 

In this blog, we will dive into a publication and a preprint that address the first two bullets.   

Urine not gonna believe this: immune checkpoint inhibitor therapy-induced signature of kidney injury 

Immune checkpoint inhibitor (ICI) therapy is a cornerstone treatment for many cancers, but as many as 60% of patients receiving ICI develop immune-related adverse events (irAEs). On one hand, irAEs are typically associated with a response to therapy, but they can also be acutely severe. One out of five patients with ICI-associated irAEs report acute kidney injury (AKI), while only 2–5% are specifically diagnosed with acute interstitial nephritis (AIN). AKIs result in the temporary suspension of ICI therapy to determine the cause, and invasive biopsies are required for AIN diagnosis (see figure). Not only can this disruption delay the reintroduction of therapy by up to four weeks, but patients are also subjected to further invasive biopsies that can cause excessive bleeding. Furthermore, delays in diagnosis put over 50% of patients at risk of permanent renal damage. The need for a non-invasive and rapid diagnostic for AIN cannot be overstated. 

To address this need, Jamie Lin and colleagues at the MD Anderson Cancer Center turned to the NULISAseq Inflammation 250 Panel to identify low-abundance markers associated with AIN in patient urine and plasma. “We’ve been waiting for an assay with this sensitivity,” Dr. Lin said in a recent webinar on immuno-oncology. “This is really a big breakthrough for urine analysis,” she added about the NULISA platform.  

The team found that urine proteomics outperformed plasma results in distinguishing ICI-AIN from non-AIN cases (Long et al., 2025). In the urine biomarker analysis, they identified 10 key proteins, two of which confirmed previous findings (CXCL9 and TNF-⍺), while eight were novel (IL-5, Fas, TNFSF4, CD274, IL-20, TNFSF15, TSLP, TREM1, and CCL1). The careful curation and multiplexing of important immune-related markers in the panel thus provided a previously undescribed signature and instilled confidence in the results, effectively recapitulating the literature.  

Using machine learning and computational approaches, the team refined their signature to two proteins—IL-5 and Fas—that achieved an area under the curve of 0.94, indicating a remarkably sensitive and selective diagnosis of ICI-AIN. Notably, these two markers were complementary in sensitivity and selectivity, with IL-5 showing 100% sensitivity and Fas displaying 100% specificity.  

With further validation, the two-protein signature could eventually become a bedside uremic diagnostic tool for rapidly identifying patients receiving ICI who likely require glucocorticoid co-therapy to alleviate AIN. 

“I’m personally excited to see where the NULISA platform is going to take us. We expect to see big changes in the nephrology world with this assay,” Dr. Lin concluded in her webinar.  

Figure: Current (left, red box) and proposed (right, yellow box) standard of care in identifying and treating acure kidney injury (AKI) in patients receiving immune checkpoint inhibitor (ICI) therapy. Adopted from Dr. Jamie Lin’s webinar on April 30, 2025. 

Let’s tox about markers of immune toxicity in CAR-T cell therapy 

CAR-T therapy is another application of cancer immunotherapy in dire need of irAE biomarkers. Like ICI, irAEs are a hallmark of CAR-T infusion and are indicative of patient response. Severe reactions often manifest as neurotoxicity (ICANS) and cytokine release syndrome (CRS), which can occur independently or in tandem. Symptoms of CRS are typically evident within a week after conditioning and CAR-T cell infusion, while the onset of ICANS occurs later, after effector immune cell infiltration of the blood-brain barrier (BBB) (Morris et al., 2022). The lack of sensitive techniques for exploring early-stage inflammatory changes and associated studies means that the underlying contributions to immune activation, BBB collapse, and response severity are poorly understood. 

A collaborative study between the Cleveland Clinic and Alamar Biosciences uncovered important signals of irAEs during the administration of anti-CD19 CAR-T cell therapy. In the most comprehensive characterization of immune response to CAR T-cell therapy to date, the team utilized the NULISAseq Inflammation Panel 250 in a longitudinal cohort of 80 patients with B-cell lymphoma (LBCL), involving 480 plasma samples over six time points (baseline and up to 16 days post-anti-CD19 CAR-T cell infusion) and including ASTCT consensus criteria scores for ICANS and CRS.  

The data read like a contour map, displaying the peaks and valleys of effector immune cell activation, exhaustion, and resolution pathways (Kirkpatrick et al., 2024). The superior sensitivity of the NULISA platform is on constant display, revealing novel markers previously unreported in CAR-T immune response, including several chemokines (CX3CL1, CXCL1, CCL4, CXCL10), Th2 T-cell effector cytokines (IL-4, IL-5, IL-13), and T-cell exhaustion proteins (CTLA4, PDCD1, CD274, LAG3). These results also confirm the known role of IL-17 in mediating immune injury, which is implicated in permeabilizing the BBB (Rahman et al., 2018). Over days 1–9, this crescendo was notably more profound in the severe irAE groups. In contrast, the irAE groups showed downregulated markers of growth and repair mediators, including PDGFA, EGF, and brain-derived neurotrophic factor (BDNF), limited access to the latter of which may worsen ICANS.  

Overall, this tour-de-force study opens the door to a better understanding of the causative mechanisms of acute immune injury in CAR-T cell therapy and informs several mitigation strategies pending further validation. We eagerly await the publication of this exciting preprint.  

Automated, ultra-high sensitivity with comprehensive coverage 

In both RNA with Advanced Cell Diagnostics and protein detection with Alamar Biosciences, Dr. Luo recognized that the key to unlocking the potential of measuring these biomolecules lies in enhancing the signal-to-noise ratio by suppressing background interference. These innovations are essential in oncology, both for detecting single RNA molecules at the single-cell level (ACD) and for achieving unprecedented sensitivity and dynamic range for proteins in biofluids (Alamar). In just two short years, equipping researchers with the NULISA platform’s best-in-class biofluid protein sensitivity has paid dividends for the early detection of immune-related adverse events associated with cancer immunotherapy.  

Be sure to tune into our next blog, where we will change gears to focus on how such breadth and depth are impacting traumatic brain injury classification and biology in both humans and mouse models.